Updated on 2025/07/16

写真a

 
OHTSUKI Takashi
 
Organization
Faculty of Interdisciplinary Science and Engineering in Health Systems Professor
Position
Professor
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Degree

  • 学士 ( 東京工業大学 )

  • 修士 ( 東京大学 )

  • Doctor(Engineering) ( The University of Tokyo )

Research Interests

  • アポトーシス

  • miRNA

  • 光制御

  • RNAキャリア

  • 超音波

  • エンドソーム脱出

  • ナノキャリア

  • 拡張翻訳系

  • tRNA

  • EF-Tu

  • RNA delivery

  • siRNA

  • 光増感剤

  • RNA工学

  • RNAi

  • ケージド化合物

  • タンパク質生合成系

  • 非天然アミノ酸

  • mRNA

  • tRNA

  • siRNA

  • Biochemistry

  • Molecular Biology

  • translation

  • nonnatural amino acid

  • RNAi

  • EF-Tu

  • messenger RNA

Research Areas

  • Nanotechnology/Materials / Bio chemistry

  • Life Science / Structural biochemistry

  • Manufacturing Technology (Mechanical Engineering, Electrical and Electronic Engineering, Chemical Engineering) / Biofunction and bioprocess engineering

  • Nanotechnology/Materials / Chemistry and chemical methodology of biomolecules

  • Life Science / Biomedical engineering

  • Life Science / Biomaterials

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Education

  • The University of Tokyo   工学研究科   化学生命工学専攻 博士課程

    1995 - 1998

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  • The University of Tokyo   工学研究科   化学生命工学専攻 修士課程

    1993 - 1995

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    Country: Japan

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  • Tokyo Institute of Technology   工学部   生体分子工学科

    1989 - 1993

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    Country: Japan

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Research History

  • Okayama University   Graduate School of Interdisciplinary Science and Engineering in Health Systems   Professor

    2018.4

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  • 岡山大学自然科学研究科 教授

    2010.6 - 2018.3

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  • 岡山大学自然科学研究科 准教授

    2007.10 - 2010.5

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  • Okayama University   Faculty of Engineering, Department of Bioscience and Biotechnology

    2003.11 - 2007.9

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  • The University of Tokyo   Graduate School of Frontier Sciences

    2002.4 - 2003.10

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  • The University of Tokyo   The Graduate School of Engineering

    2000.3 - 2002.3

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  • 理化学研究所ゲノム化学総合研究センター リサーチアソシエイト

    1999.1 - 2000.3

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  • 日本学術振興会 特別研究員 日本学術振興会特別研究員

    1997.4 - 1998.12

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Professional Memberships

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Committee Memberships

  • 日本光医学・光生物学会   評議員  

    2023.4   

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    Committee type:Academic society

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  • 日本化学会中国四国支部   役員  

    2021.4 - 2023.3   

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  • 日本核酸化学会   評議員  

    2016   

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    Committee type:Academic society

    日本核酸化学会

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  • 日本生化学会   代議員  

    2014.4 - 2018.3   

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    Committee type:Academic society

    日本生化学会

  • 日本化学会 生体機能関連化学部会   ニュースレター編集委員  

    2011.4 - 2014.3   

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    Committee type:Academic society

    日本化学会 生体機能関連化学部会

  • 日本生化学会   評議員  

    2011   

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    Committee type:Academic society

    日本生化学会

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  • 日本化学会 生体機能関連化学部会   幹事  

    2010   

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    Committee type:Academic society

    日本化学会 生体機能関連化学部会

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Papers

  • Photochemical internalization of mRNA using a photosensitizer and nucleic acid carriers Reviewed

    Hayaki Maemoto, Ryohei Suzaki, Kazunori Watanabe, Keiji Itaka, Takashi Ohtsuki

    European Journal of Medicinal Chemistry Reports   13   100242 - 100242   2025.4

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    Authorship:Last author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.ejmcr.2024.100242

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  • Novel Drug Delivery Particles Can Provide Dual Effects on Cancer “Theranostics” in Boron Neutron Capture Therapy Reviewed

    Abdul Basith Fithroni, Haruki Inoue, Shengli Zhou, Taufik Fatwa Nur Hakim, Takashi Tada, Minoru Suzuki, Yoshinori Sakurai, Manabu Ishimoto, Naoyuki Yamada, Rani Sauriasari, Wolfgang A. G. Sauerwein, Kazunori Watanabe, Takashi Ohtsuki, Eiji Matsuura

    Cells   14 ( 1 )   60 - 60   2025.1

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    Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    Boron (B) neutron capture therapy (BNCT) is a novel non-invasive targeted cancer therapy based on the nuclear capture reaction 10B (n, alpha) 7Li that enables the death of cancer cells without damaging neighboring normal cells. However, the development of clinically approved boron drugs remains challenging. We have previously reported on self-forming nanoparticles for drug delivery consisting of a biodegradable polymer, namely, “AB-type” Lactosome® nanoparticles (AB-Lac particles)- highly loaded with hydrophobic B compounds, namely o-Carborane (Carb) or 1,2-dihexyl-o-Carborane (diC6-Carb), and the latter (diC6-Carb) especially showed the “molecular glue” effect. Here we present in vivo and ex vivo studies with human pancreatic cancer (AsPC-1) cells to find therapeutically optimal formulas and the appropriate treatment conditions for these particles. The biodistribution of the particles was assessed by the tumor/normal tissue ratio (T/N) in terms of tumor/muscle (T/M) and tumor/blood (T/B) ratios using near-infrared fluorescence (NIRF) imaging with indocyanine green (ICG). The in vivo and ex vivo accumulation of B delivered by the injected AB-Lac particles in tumor lesions reached a maximum by 12 h post-injection. Irradiation studies conducted both in vitro and in vivo showed that AB-Lac particles-loaded with either 10B-Carb or 10B-diC6-Carb significantly inhibited the growth of AsPC-1 cancer cells or strongly inhibited their growth, with the latter method being significantly more effective. Surprisingly, a similar in vitro and in vivo irradiation study showed that ICG-labeled AB-Lac particles alone, i.e., without any 10B compounds, also revealed a significant inhibition. Therefore, we expect that our ICG-labeled AB-Lac particles-loaded with 10B compound(s) may be a novel and promising candidate for providing not only NIRF imaging for a practical diagnosis but also the dual therapeutic effects of induced cancer cell death, i.e., “theranostics”.

    DOI: 10.3390/cells14010060

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  • Configuration of two cysteine residues in a ring within a stapled Bim peptide affects the secondary structure and apoptotic activity Reviewed

    Shengli Zhou, Fuka Nishimura, Kazuhaya Wada, Kaho Fujii, Takeshi Kondo, Kazunori Watanabe, Yoshitane Imai, Takashi Ohtsuki, Mizuki Kitamatsu

    Bioorganic & Medicinal Chemistry Letters   112   129915 - 129915   2024.11

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    DOI: 10.1016/j.bmcl.2024.129915

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  • In Vitro Study of Tumor-Homing Peptide-Modified Magnetic Nanoparticles for Magnetic Hyperthermia Reviewed

    Shengli Zhou, Kaname Tsutsumiuchi, Ritsuko Imai, Yukiko Miki, Anna Kondo, Hiroshi Nakagawa, Kazunori Watanabe, Takashi Ohtsuki

    Molecules   29 ( 11 )   2632 - 2632   2024.6

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    Cancer cells have higher heat sensitivity compared to normal cells; therefore, hyperthermia is a promising approach for cancer therapy because of its ability to selectively kill cancer cells by heating them. However, the specific and rapid heating of tumor tissues remains challenging. This study investigated the potential of magnetic nanoparticles (MNPs) modified with tumor-homing peptides (THPs), specifically PL1 and PL3, for tumor-specific magnetic hyperthermia therapy. The synthesis of THP-modified MNPs involved the attachment of PL1 and PL3 peptides to the surface of the MNPs, which facilitated enhanced tumor cell binding and internalization. Cell specificity studies revealed an increased uptake of PL1- and PL3-MNPs by tumor cells compared to unmodified MNPs, indicating their potential for targeted delivery. In vitro hyperthermia experiments demonstrated the efficacy of PL3-MNPs in inducing tumor cell death when exposed to an alternating magnetic field (AMF). Even without exposure to an AMF, an additional ferroptotic pathway was suggested to be mediated by the nanoparticles. Thus, this study suggests that THP-modified MNPs, particularly PL3-MNPs, hold promise as a targeted approach for tumor-specific magnetic hyperthermia therapy.

    DOI: 10.3390/molecules29112632

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  • Pyrene-Modified Cyclic Peptides Detect Cu2+ Ions by Fluorescence in Water Reviewed

    Yuhi Maekawa, Sora Sakura, Yuji Furutani, Rento Fujihara, Hisashi Sugime, Takashi Ohtsuki, Mizuki Kitamatsu

    Processes   12 ( 4 )   746 - 746   2024.4

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    The detection of metal ions is an option for maintaining water quality and diagnosing metal ion-related diseases. In this study, we successfully detected metal ions using fluorescent peptides in water. First, we prepared seven linear (L1–L7) and seven cyclic (C1–C7) peptides containing two pyrenyl (Pyr) units and assessed the response to various metal ions by fluorescence. The results indicated that C1, which contains a hexameric cyclic peptide moiety consisting of Pyr and Gly units, did not show a fluorescent response to metal ions, while the linear L1 corresponding to C1 showed a response to Cu2+, but its selectivity was found to be poor through a competition assay for each metal ion. We then assessed C2–C7 and L2–L7, in which Gly was replaced by His units at various positions in the same manner. The results showed that C2–C7 responded to Cu2+ in a manner dependent on the His position. Additionally, superior selectivity was observed in C7 through a competition assay. These results demonstrate that the structural restriction of peptides and the sequence affect the selective detection of Cu2+ and reveal that peptides with an appropriate structure can accomplish the fluorescent detection of Cu2+ specifically.

    DOI: 10.3390/pr12040746

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  • Self-assembly of Peptide Amphiphiles with Alkyl Groups for siRNA Delivery Reviewed

    Taufik F.N. Hakim, Kazunori Watanabe, Shomu Fujimoto, Mizuki Kitamatsu, Takashi Ohtsuki

    Chemistry Letters   2023.9

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    Authorship:Last author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:The Chemical Society of Japan  

    DOI: 10.1246/cl.230302

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  • Photo-dependent cytosolic delivery of shRNA into a single blastomere in a mouse embryo. Reviewed International journal

    Yuka Ikawa, Takuya Wakai, Hiroaki Funahashi, Tet Htut Soe, Kazunori Watanabe, Takashi Ohtsuki

    Scientific reports   13 ( 1 )   13050 - 13050   2023.8

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    Single-cell-specific delivery of small RNAs, such as short hairpin RNA (shRNA) and small noncoding RNAs, allows us to elucidate the roles of specific upregulation of RNA expression and RNAi-mediated gene suppression in early embryo development. The photoinduced cytosolic dispersion of RNA (PCDR) method that we previously reported can introduce small RNAs into the cytosol of photoirradiated cells and enable RNA delivery into a single-cell in a spatiotemporally specific manner. However, the PCDR method has only been applied to planer cultured cells and not to embryos. This study demonstrated that the PCDR method can be utilized for photo-dependent cytosolic shRNA delivery into a single blastomere and for single blastomere-specific RNA interference in mouse embryos. Our results indicate that PCDR is a promising approach for studying the developmental process of early embryogenesis.

    DOI: 10.1038/s41598-023-40361-9

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  • FRET probe for detecting two mutations in one EGFR mRNA. Reviewed International journal

    Myat Thu, Kouta Yanai, Hajime Shigeto, Shohei Yamamura, Kazunori Watanabe, Takashi Ohtsuki

    Analyst   148 ( 11 )   2626 - 2632   2023.5

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    Technologies for visualizing and tracking RNA are essential in molecular biology, including in disease-related fields. In this study, we propose a novel probe set (DAt-probe and T-probe) that simultaneously detects two mutations in the same RNA using fluorescence resonance energy transfer (FRET). The DAt-probe carrying the fluorophore Atto488 and the quencher Dabcyl were used to detect a cancer mutation (exon19del), and the T-probe carrying the fluorophore Tamra was used to detect drug resistance mutations (T790M) in epidermal growth factor receptor (EGFR) mRNA. These probes were designed to induce FRET when both mutations were present in the mRNA. Gel electrophoresis confirmed that the two probes could efficiently bind to the mutant mRNA. We measured the FRET ratios using wild-type and double-mutant RNAs and found a significant difference between them. Even in living cells, the FRET probe could visualize mutant RNA. As a result, we conclude that this probe set provides a method for detecting two mutations in the single EGFR mRNA via FRET.

    DOI: 10.1039/d3an00554b

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  • Photochemical internalizationに基づく光依存的細胞質内RNA導入法の副作用低減 Reviewed

    Akinari Bando, Kazunori Watanabe, Takashi Ohtsuki

    The Journal of Japan Society for Laser Surgery and Medicine   44 ( 1 )   62 - 68   2023.4

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    Authorship:Last author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Japan Society for Laser Surgery and Medicine  

    DOI: 10.2530/jslsm.jslsm-44_0004

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  • Reduction of the side effects of photoinduced cytosolic dispersion of RNA (PCDR) based on photochemical internalization Invited Reviewed International journal

    Akinari Bando, Kazunori Watanabe, Takashi Ohtsuki

    2023.4

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    Authorship:Last author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)  

    BNCT is a non-invasive cancer therapy that allows for cancer cell death without harming adjacent cells. However, the application is limited, owing to the challenges of working with clinically approved boron (B) compounds and drug delivery systems (DDS). To address the issues, we developed self-forming nanoparticles consisting of a biodegradable polymer, namely, "AB-type Lactosome (AB-Lac)" loaded with B compounds. Three carborane isomers (o-, m-, and p-carborane) and three related alkylated derivatives, i.e., 1,2-dimethy-o-carborane (diC1-Carb), 1,2-dihexyl-o-carborane (diC6-Carb), and 1,2-didodecyl-o-carborane (diC12-Carb), were separately loaded. diC6-Carb was highly loaded with AB-Lac particles, and their stability indicated the "molecular glue" effect. The efficiency of in vitro B uptake of diC6-Carb for BNCT was confirmed at non-cytotoxic concentration in several cancer cell lines. In vivo/ex vivo biodistribution studies indicated that the AB-Lac particles were remarkably accumulated within 72 h post-injection in the tumor lesions of mice bearing syngeneic breast cancer (4T1) cells, but the maximum accumulation was reached at 12 h. In ex vivo B biodistribution, the ratios of tumor/normal tissue (T/N) and tumor/blood (T/Bl) of the diC6-Carb-loaded particles remained stably high up to 72 h. Therefore, we propose the diC6-Carb-loaded AB-Lac particles as a promising candidate medicine for BNCT.

  • Adjusting Heterodimeric Coiled-Coils (K/E Zipper) to Connect Autophagy-Inducing Peptide with Cell-Penetrating Peptide. Reviewed International journal

    Yoshiyuki Hakata, Kazuma Yamashita, Sonoko Hashimoto, Takashi Ohtsuki, Masaaki Miyazawa, Mizuki Kitamatsu

    Pharmaceutics   15 ( 4 )   2023.3

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    DOI: 10.3390/pharmaceutics15041048

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  • Novel Self-Forming Nanosized DDS Particles for BNCT: Utilizing A Hydrophobic Boron Cluster and Its Molecular Glue Effect. Reviewed International journal

    Abdul Basith Fithroni, Kazuko Kobayashi, Hirotaka Uji, Manabu Ishimoto, Masaru Akehi, Takashi Ohtsuki, Eiji Matsuura

    Cells   11 ( 20 )   2022.10

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    DOI: 10.3390/cells11203307

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  • Fluorescence ratiometric DNA detection by peptide nucleic acid-pyrene binary probes. Reviewed International journal

    Koki Ishii, Sakura Tsuchitani, Miyu Toyama, Hajime Shigeto, Shohei Yamamura, Takashi Ohtsuki, Yoshitane Imai, Mizuki Kitamatsu

    Bioorganic & medicinal chemistry letters   71   128838 - 128838   2022.9

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    DOI: 10.1016/j.bmcl.2022.128838

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  • Ultrasound-dependent RNAi using TatU1A-rose bengal conjugate. Reviewed International journal

    Nanako Sumi, Shota Nagahiro, Eiji Nakata, Kazunori Watanabe, Takashi Ohtsuki

    Bioorganic & medicinal chemistry letters   68   128767 - 128767   2022.7

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    Tat-U1A-rose bengal conjugate (TatU1A-RB) was prepared as an ultrasound-sensitive RNA carrier molecule. This molecule consists of Tat cell-penetrating peptide, U1A RNA-binding protein, and rose bengal as a sonosensitizer. We demonstrated that TatU1A-RB delivered RNA via the endocytosis pathway, which was followed by ultrasound-dependent endosomal escape and cytosolic dispersion of the RNA. A short hairpin RNA (shRNA) delivered by TatU1A-RB mediated RNA interference (RNAi) ultrasound-dependently. Even by ultrasound irradiation through blood cells, RNAi could be induced with TatU1A-RB and the shRNA. This ultrasound-dependent cytosolic RNA delivery method will serve as the basis for a new approach to nucleic acid therapeutics.

    DOI: 10.1016/j.bmcl.2022.128767

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  • Adjusting the Structure of a Peptide Nucleic Acid (PNA) Molecular Beacon and Promoting Its DNA Detection by a Hybrid with Quencher-Modified DNA Reviewed

    Hajime Shigeto, Takashi Ohtsuki, Shohei Yamamura, Mizuki Kitamatsu

    PROCESSES   10 ( 4 )   2022.4

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    DOI: 10.3390/pr10040722

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  • Fluorophore-PNA-Quencher/Quencher-DNA probe for miRNA detection. Reviewed International journal

    Kentaro Tabara, Kazunori Watanabe, Hajime Shigeto, Shohei Yamamura, Takamasa Kishi, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   51   128359 - 128359   2021.9

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    Micro RNAs (miRNAs) are involved in a variety of biological functions and are attracting attention as diagnostic and prognostic markers for various diseases. Highly sensitive RNA detection methods are required to determine miRNA expression levels and intracellular localization. In this study, we designed new double-stranded peptide nucleic acid (PNA)/DNA probes consisting of a fluorophore-PNA-quencher (fPq) and a quencher-DNA (qD) for miR-221 detection. We optimized the fPq structure, PNA-DNA hybrid length, and hybrid position. The resultant fPq-2/qD-6b probe was a 6-bp hybrid probe with a 10-base fPq and a 6-base qD. The signal-to-background ratios of the probes showed that fPq-2/qD-6b had a higher target sensitivity than fPq (PNA beacon)-type and fP/qD-type probes. The results of the detection limit and target specificity indicate that the fPq/qD probe is promising for RNA detection in both cells and cell extracts as well as for miRNA diagnosis.

    DOI: 10.1016/j.bmcl.2021.128359

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  • Photocontrolled apoptosis induction using precursor miR-664a and an RNA carrier-conjugated with photosensitizer. Reviewed International journal

    Kazunori Watanabe, Tomoko Nawachi, Ruriko Okutani, Takashi Ohtsuki

    Scientific reports   11 ( 1 )   14936 - 14936   2021.7

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    DOI: 10.1038/s41598-021-94249-7.

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  • Nucleic Acids Chemistry and Engineering: Special Issue on Nucleic Acid Conjugates for Biotechnological Applications

    Tamaki Endoh, Eriks Rozners, Takashi Ohtsuki

    Applied Sciences   11 ( 8 )   2021.4

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  • Lactosome-Conjugated siRNA Nanoparticles for Photo-Enhanced Gene Silencing in Cancer Cells. Reviewed International journal

    Melissa Siaw Han Lim, Yuki Nishiyama, Takashi Ohtsuki, Kazunori Watanabe, Hirotsugu Kobuchi, Kazuko Kobayashi, Eiji Matsuura

    Journal of pharmaceutical sciences   110 ( 4 )   1788 - 1798   2021.4

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.xphs.2021.01.026

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  • Minimization of apoptosis-inducing CPP-Bim peptide. Reviewed International journal

    Shengli Zhou, Kazunori Watanabe, Seiichiro Koide, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & medicinal chemistry letters   36   127811 - 127811   2021.3

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    DOI: 10.1016/j.bmcl.2021.127811

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  • Inhibition of HSF1 and SAFB Granule Formation Enhances Apoptosis Induced by Heat Stress Reviewed

    Watanabe K, Ohtsuki T.

    Int J Mol Sci.   22 ( 9 )   4982 - 4982   2021.3

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    DOI: 10.3390/ijms22094982.

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  • A Novel 89Zr-labeled DDS Device Utilizing Human IgG Variant (scFv): "Lactosome" Nanoparticle-Based Theranostics for PET Imaging and Targeted Therapy. Reviewed International journal

    Melissa Siaw Han Lim, Takashi Ohtsuki, Fumiaki Takenaka, Kazuko Kobayashi, Masaru Akehi, Hirotaka Uji, Hirotsugu Kobuchi, Takanori Sasaki, Eiichi Ozeki, Eiji Matsuura

    Life   11 ( 2 )   2021.2

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    "Theranostics," a new concept of medical advances featuring a fusion of therapeutic and diagnostic systems, provides promising prospects in personalized medicine, especially cancer. The theranostics system comprises a novel 89Zr-labeled drug delivery system (DDS), derived from the novel biodegradable polymeric micelle, "Lactosome" nanoparticles conjugated with specific shortened IgG variant, and aims to successfully deliver therapeutically effective molecules, such as the apoptosis-inducing small interfering RNA (siRNA) intracellularly while offering simultaneous tumor visualization via PET imaging. A 27 kDa-human single chain variable fragment (scFv) of IgG to establish clinically applicable PET imaging and theranostics in cancer medicine was fabricated to target mesothelin (MSLN), a 40 kDa-differentiation-related cell surface glycoprotein antigen, which is frequently and highly expressed by malignant tumors. This system coupled with the cell penetrating peptide (CPP)-modified and photosensitizer (e.g., 5, 10, 15, 20-tetrakis (4-aminophenyl) porphyrin (TPP))-loaded Lactosome particles for photochemical internalized (PCI) driven intracellular siRNA delivery and the combination of 5-aminolevulinic acid (ALA) photodynamic therapy (PDT) offers a promising nano-theranostic-based cancer therapy via its targeted apoptosis-inducing feature. This review focuses on the combined advances in nanotechnology and material sciences utilizing the "89Zr-labeled CPP and TPP-loaded Lactosome particles" and future directions based on important milestones and recent developments in this platform.

    DOI: 10.3390/life11020158

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  • Complementary leucine zippering system for effective intracellular delivery of proteins by cell-penetrating peptides. Reviewed International journal

    Mizuki Kitamatsu, Hiroki Yuasa, Takashi Ohtsuki, Hiroyuki Michiue

    Bioorganic & medicinal chemistry   33   116036 - 116036   2021.1

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    DOI: 10.1016/j.bmc.2021.116036

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  • Fluorescence lifetime probes for detection of RNA degradation Reviewed International journal

    Riku Hirata, Kazutaka Hirakawa, Naotaka Shimada, Kazunori Watanabe, Takashi Ohtsuki

    Analyst   146 ( 1 )   277 - 282   2021.1

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    DOI: 10.1039/d0an01230k

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  • Photoinduced Endosomal Escape Mechanism: A View from Photochemical Internalization Mediated by CPP-Photosensitizer Conjugates. Invited Reviewed International journal

    Tet Htut Soe, Kazunori Watanabe, Takashi Ohtsuki

    Molecules   26 ( 1 )   2020.12

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.3390/molecules26010036

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  • Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate. Reviewed International journal

    Hyungjin Kim, Sho Watanabe, Mizuki Kitamatsu, Kazunori Watanabe, Takashi Ohtsuki

    Scientific reports   10 ( 1 )   19087 - 19087   2020.11

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/s41598-020-76100-7

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  • Analysis of Single Nucleotide-Mutated Single-Cancer Cells Using the Combined Technologies of Single-Cell Microarray Chips and Peptide Nucleic Acid-DNA Probes. Reviewed International journal

    Hajime Shigeto, Eriko Yamada, Mizuki Kitamatsu, Takashi Ohtsuki, Akira Iizuka, Yasuto Akiyama, Shohei Yamamura

    Micromachines   11 ( 7 )   E628 - E628   2020.6

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    Research into cancer cells that harbor gene mutations relating to anticancer drug-resistance at the single-cell level has focused on the diagnosis of, or treatment for, cancer. Several methods have been reported for detecting gene-mutated cells within a large number of non-mutated cells; however, target single nucleotide-mutated cells within a large number of cell samples, such as cancer tissue, are still difficult to analyze. In this study, a new system is developed to detect and isolate single-cancer cells expressing the T790M-mutated epidermal growth factor receptor (EGFR) mRNA from multiple non-mutated cancer cells by combining single-cell microarray chips and peptide nucleic acid (PNA)-DNA probes. The single-cell microarray chip is made of polystyrene with 62,410 microchambers (31-40 µm diameter). The T790M-mutated lung cancer cell line, NCI-H1975, and non-mutated lung cancer cell line, A549, were successfully separated into single cells in each microchambers on the chip. Only NCI-H1975 cell was stained on the chip with a fluorescein isothiocyanate (FITC)-conjugated PNA probe for specifically detecting T790M mutation. Of the NCI-H1975 cells that spiked into A549 cells, 0-20% were quantitatively analyzed within 1 h, depending on the spike concentration. Therefore, our system could be useful in analyzing cancer tissue that contains a few anticancer drug-resistant cells.

    DOI: 10.3390/mi11070628

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  • Endosomal Escape of Peptide-Photosensitizer Conjugates Is Affected by Amino Acid Sequences near the Photosensitizer. Reviewed International journal

    Yuichi Miyoshi, Maho Kadono, Shigetoshi Okazaki, Ayano Nishimura, Mizuki Kitamatsu, Kazunori Watanabe, Takashi Ohtsuki

    Bioconjugate Chemistry   31 ( 3 )   916 - 922   2020.3

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    DOI: 10.1021/acs.bioconjchem.0c00046

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  • Intracellular delivery of a peptide nucleic acid-based hybrid of an autophagy inducing peptide with a cell-penetrating peptide Reviewed

    Yoshiyuki Hakata, Suzuka Ishikawa, Takashi Ohtsuki, Masaaki Miyazawa, Mizuki Kitamatsu

    Organic & Biomolecular Chemistry   18 ( 10 )   1978 - 1986   2020.3

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  • Relation of Photochemical Internalization to Heat, pH and Ca2+ Ions. Reviewed International journal

    Tet Htut Soe, Tomotaka Nanjo, Kazunori Watanabe, Takashi Ohtsuki

    Photochemistry and photobiology   95 ( 6 )   1395 - 1402   2019.11

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    The inefficient endosomal escape of drugs or macromolecules is a major obstacle to achieving successful delivery to therapeutic targets. An efficient approach to circumvent this barrier is photochemical internalization (PCI), which uses light and photosensitizers for endosomal escape of the delivered macromolecules. The PCI mechanism is related to photogenerated singlet oxygen, but the mechanism is still unclear. In this study, we examined the relation of PCI to heat, pH and Ca2+ ions using cell penetrating peptide (CPP)-cargo-photosensitizer (Alexa546 or Alexa633) conjugates. A cell temperature changing experiment demonstrated that heat (thermal mechanism) does not significantly contribute to the photoinduced endosomal escape. Inhibition of V-ATPase proton pump activity and endosomal pH upregulation indicated that PCI-mediated endosomal escape needs endosomal acidification prior to photoirradiation. Imaging of the CPP-cargo-photosensitizer and Ca2+ ions during photostimulation showed that intracellular calcium increase is not the cause of the endosomal escape of the complex. The increment is mainly due to Ca2+ influx. These findings show the importance of extra- and intracellular milieu conditions in the PCI mechanism and enrich our understanding of PCI-related changes in cell.

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  • Imaging analysis of EGFR mutated cancer cells using peptide nucleic acid (PNA)-DNA probes. Reviewed

    Shigeto H, Ohtsuki T, Iizuka A, Akiyama Y, Yamamura S

    Analyst   144 ( 15 )   4613 - 4621   2019.8

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  • A leucine zipper-based peptide hybrid delivers functional Nanog protein inside the cell nucleus. Reviewed International journal

    Yoshiyuki Hakata, Hiroyuki Michiue, Takashi Ohtsuki, Masaaki Miyazawa, Mizuki Kitamatsu

    Bioorganic & medicinal chemistry letters   29 ( 7 )   878 - 881   2019.4

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    DOI: 10.1016/j.bmcl.2019.02.004

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  • Combined apoptotic effects of peptide and miRNA in a peptide/miRNA nanocomplex Reviewed

    Hyungjin Kim, Mizuki Kitamatsu, Takashi Ohtsuki

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   128 ( 1 )   110 - 116   2019.1

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    DOI: 10.1016/j.jbiosc.2019.01.003

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  • In-stem molecular beacon targeted to a 5'-region of tRNA inclusive of the D arm that detects mature tRNA with high sensitivity. Reviewed International journal

    Miyoshi Y, Ohtsuki T, Kashida H, Asanuma H, Watanabe K

    PloS one   14 ( 1 )   e0211505   2019

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  • Red and Near-Infrared Light-Directed Cytosolic Delivery of Two Different RNAs Using Photosensitive RNA Carriers. Reviewed International journal

    Kaori Shiraga, Tet Htut Soe, Sho Matsumoto, Kazunori Watanabe, Takashi Ohtsuki

    Bioconjugate chemistry   29 ( 9 )   3174 - 3179   2018.8

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    Many cellular events are thought to be controlled by the temporal upregulation of multiple RNAs; the timing of the upregulation of these RNAs is not always the same. In this study, we first show that our light-directed intracellular RNA delivery method induced high concentrations of RNA in a short period. This effect was beneficial for the temporal control of cellular events by functional RNAs. Next, we stimulated the short-term upregulation of two different RNAs at different time points. Cytosolic delivery of a first RNA was induced by red light; thereafter, cytosolic delivery of a second RNA was induced by near-infrared light. The time difference between the introduction of the first and second RNA can be short (0.5-4 h) or long (>8 h). This strategy shows the potential for future applications of the deliberate control of time-dependent RNA concentration to guide various cellular functions by multiple RNAs.

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  • MicroRNA-664a-5p promotes neuronal differentiation of SH-SY5Y cells Reviewed

    Kazunori Watanabe, Ryuhei Yamaji, Takashi Ohtsuki

    Genes to Cells   23 ( 3 )   225 - 233   2018.3

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    DOI: 10.1111/gtc.12559

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  • Enhanced intracellular peptide delivery by multivalent cell-penetrating peptide with bioreducible linkage Reviewed

    Hyungjin Kim, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   28 ( 3 )   378 - 381   2018.2

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    DOI: 10.1016/j.bmcl.2017.12.035

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  • Ultrasound-dependent cytoplasmic internalization of a peptidesonosensitizer conjugate Reviewed

    Yuki Inaba, Kazunori Watanabe, Mizuki Kitamatsu, Eiji Nakata, Atsushi Harada, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry   25 ( 15 )   4212 - 4217   2017.8

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    DOI: 10.1016/j.bmc.2017.06.024

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  • Detection of small, highly structured RNAs using molecular beacons Reviewed

    J. Li, C. Xu, N. Shimada, Y. Miyoshi, K. Watanabe, W. Cong, T. Ohtsuki

    Analytical Methods   9 ( 20 )   2971 - 2976   2017.5

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    DOI: 10.1039/c7ay00341b

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  • Duplication of Drosophila melanogaster mitochondrial EF-Tu: pre-adaptation to T-arm truncation and exclusion of bulky aminoacyl residues Reviewed

    Aya Sato, Takuma Suematsu, Koh-Ki Aihara, Kiyoshi Kita, Tsutomu Suzuki, Kimitsuna Watanabe, Takashi Ohtsuki, Yoh-ichi Watanabe

    Biochemical Journal   474   957 - 969   2017.3

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    DOI: 10.1042/BCJ20160929

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  • Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs Reviewed

    Takashi Ohtsuki, Shigeto Kanzaki, Sae Nishimura, Yoshio Kunihiro, Masahiko Sisido, Kazunori Watanabe

    Nature Communications   7   12501 - 12501   2016.8

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    DOI: 10.1038/ncomms12501

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  • Photoinduced apoptosis using a peptide carrying a photosensitizer Reviewed

    Kazunori Watanabe, Hayato Fujiwara, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   26 ( 13 )   3115 - 3118   2016.7

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    DOI: 10.1016/j.bmcl.2016.04.091

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  • Enhanced cellular uptake of lactosomes using cell-penetrating peptides. Reviewed International journal

    Akiya Akahoshi, Eiji Matsuura, Eiichi Ozeki, Hayato Matsui, Kazunori Watanabe, Takashi Ohtsuki

    Science and technology of advanced materials   17 ( 1 )   245 - 252   2016.6

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    DOI: 10.1080/14686996.2016.1178056

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  • Photocontrolled intracellular RNA delivery by using nanoparticles or carrier-photosensitizer conjugates Invited Reviewed International journal

    Kazunori Watanabe, Takashi Ohtsuki

    Prog Mol Biol Transl Sci.   139   101 - 119   2016

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    DOI: 10.1016/bs.pmbts.2015.10.013

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  • The molecular mechanism of photochemical internalization of cell penetrating peptide-cargophoto-sensitizer conjugates Reviewed

    Takashi Ohtsuki, Shunya Miki, Shouhei Kobayashi, Tokuko Haraguchi, Eiji Nakata, Kazutaka Hirakawa, Kensuke Sumita, Kazunori Watanabe, Shigetoshi Okazaki

    Scientific Reports   5   18577 - 18577   2015.12

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    DOI: 10.1038/srep18577

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  • A novel leucine zipper motif-based hybrid peptide delivers a functional peptide cargo inside cells Reviewed

    Hakata, Y. Tsuchiya, S, Michiue, H, Ohtsuki, T, Matsui, H, Miyazawa M, Kitamatsu, M

    Chemical Communications   51   413 - 416   2015.5

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  • Photo-dependent protein biosynthesis using a caged aminoacyl-tRNA Reviewed

    Akiya Akahoshi, Yoshio Doi, Masahiko Sisido, Kazunori Watanabe, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   24 ( 23 )   5369 - 5372   2014.12

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    DOI: 10.1016/j.bmcl.2014.10.053

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  • mTOR regulates the nucleoplasmic diffusion of Xrn2 under conditions of heat stress Reviewed

    Kazunori Watanabe, Kenichi Ijiri, Takashi Ohtsuki

    FEBS Letters   588 ( 18 )   3454 - 3460   2014.9

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    DOI: 10.1016/j.febslet.2014.08.003

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  • Losing the stem-loop structure from metazoan mitochondrial tRNAs and co-evolution of interacting factors Reviewed

    Yoh-ichi Watanabe, Takuma Suematsu, Takashi Ohtsuki

    Frontiers in Genetics   5   109   2014.5

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    DOI: 10.3389/fgene.2014.00109

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  • Intracellular Delivery of RNA via RNA-Binding Proteins or Peptides Invited Reviewed

    Kazunori Watanabe, Takashi Ohtsuki

    Intracellular Delivery II   403 - 416   2014

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    DOI: 10.1007/978-94-017-8896-0_19

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  • Near-infrared light-directed RNAi using a photosensitive carrier molecule Reviewed

    Yuka Matsushita-Ishiodori, Mika Morinaga, Kazunori Watanabe, Takashi Ohtsuki

    Bioconjugate Chemistry   24 ( 10 )   1669 - 1673   2013.10

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    DOI: 10.1021/bc4001195

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  • Synthesis and Properties of Peptide Dendrimers Containing Fluorescent and Branched Amino Acids Reviewed

    Mizuki Kitamatsu, Mayumi Kitabatake, Yoshiteru Noutoshi, Takashi Ohtsuki

    Biopolymers   100 ( 1 )   64 - 70   2013

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    DOI: 10.1002/bip.22175

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  • Photoinduced RNA Interference Invited Reviewed

    Yuka Matsushita-Ishiodori, Takashi Ohtsuki

    Accounts of Chemical Research   45 ( 7 )   1039 - 1047   2012.7

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    DOI: 10.1021/ar200227n

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  • Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes Reviewed

    Takuma Ohtsuka, Satoshi Neki, Tamotsu Kanai, Kazunari Akiyoshi, Shin-ichiro M. Nomura, Takashi Ohtsuki

    Analytical Biochemistry   418 ( 1 )   97 - 101   2011.11

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    DOI: 10.1016/j.ab.2011.06.026

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  • Photosensitizing Carrier Proteins for Photoinducible RNA Interference Reviewed

    Yuka Matsushita-Ishiodori, Rina Kuwabara, Hiroyuki Sakakoshi, Tamaki Endoh, Takashi Ohtsuki

    Bioconjugate Chemistry   22 ( 11 )   2222 - 2226   2011.11

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    DOI: 10.1021/bc200095a

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  • Site-specific incorporation of arginine analogs into proteins using arginyl-tRNA synthetase Reviewed

    Akiya Akahoshi, Yoshitaka Suzue, Mizuki Kitamatsu, Masahiko Sisido, Takashi Ohtsuki

    Biochem. Biophys. Res. Commun.   414 ( 3 )   625 - 630   2011.10

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    DOI: 10.1016/j.bbrc.2011.09.137

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  • Antisense effect of pyrrolidine-based oxy-peptide nucleic acids in Escherichia coli Reviewed

    Mizuki Kitamatsu, Shunsuke Kurami, Takashi Ohtsuki, Masahiko Sisido

    Bioorganic & Medicinal Chemistry Letters   21 ( 1 )   225 - 227   2011.1

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    DOI: 10.1016/j.bmcl.2010.11.034

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  • Synthesis of a cyclic peptide/protein using the NEXT-A reaction followed by cyclization Reviewed

    Toshimasa Hamamoto, Masahiko Sisido, Takashi Ohtsuki, Masumi Taki

    Chemical Communications   47 ( 32 )   9116 - 9118   2011

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    DOI: 10.1039/c1cc12196k

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  • Synthesis of pyrrolidine-based oxy-peptide nucleic acids carrying four types of nucleobases and their transport into cytoplasm Reviewed

    Mizuki Kitamatsu, Akiko Takahashi, Takashi Ohtsuki, Masahiko Sisido

    Tetrahedron   66 ( 51 )   9659 - 9666   2010.12

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    DOI: 10.1016/j.tet.2010.10.056

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  • Use of EF-Tu mutants for determining and improving aminoacylation efficiency and for purifying aminoacyl tRNAs with non-natural amino acids Reviewed

    Takashi Ohtsuki, Hiromichi Yamamoto, Yoshio Doi, Masahiko Sisido

    Journal of Biochemistry   148 ( 2 )   239 - 246   2010.8

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    DOI: 10.1093/jb/mvq053

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  • Lactobacillus-mediated RNA interference in nematode Reviewed

    Ai Kuwahara, Masashi Arita, Akira Kushiro, Yasuji Sakube, Masahiko Sisido, Takashi Ohtsuki

    J. Biosci. Bioeng.   109 ( 2 )   189 - 192   2010.2

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    DOI: 10.1016/j.jbiosc.2009.08.002

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  • Cellular siRNA Delivery Using TatU1A and Photo-Induced RNA Interference Invited Reviewed

    Tamaki Endoh, Takashi Ohtsuki

    RNA Interference   271 - 281   2010

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    DOI: 10.1007/978-1-60761-588-0_17

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  • Detection of Bioactive Small Molecules by Fluorescent Resonance Energy Transfer (FRET) in RNA-Protein Conjugates Reviewed

    Tamaki Endoh, Ryo Shintani, Masayasu Mie, Eiry Kobatake, Takashi Ohtsuki, Masahiko Sisido

    Bioconjugate Chemistry   20 ( 12 )   2242 - 2246   2009.12

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    DOI: 10.1021/bc9002184

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  • Spatial regulation of specific gene expression through photoactivation of RNAi Reviewed

    Tamaki Endoh, Masahiko Sisido, Takashi Ohtsuki

    Journal of Controlled Release   137 ( 3-4 )   241 - 245   2009.8

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    DOI: 10.1016/j.jconrel.2009.04.015

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  • Cellular siRNA delivery using cell-penetrating peptides modified for endosomal escape Invited Reviewed

    Tamaki Endoh, Takashi Ohtsuki

    Advanced Drug Delivery Reviews   61 ( 9 )   704 - 709   2009.7

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    DOI: 10.1016/j.addr.2009.04.005

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  • Carrier PNA for shRNA delivery into cells Reviewed

    Mizuki Kitamatsu, Takanori Kubo, Rino Matsuzaki, Tamaki Endoh, Takashi Ohtsuki, Masahiko Sisido

    Bioorganic & Medicinal Chemistry Letters   19 ( 13 )   3410 - 3413   2009.7

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    DOI: 10.1016/j.bmcl.2009.05.031

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  • A protease inhibitor discovery method using fluorescence correlation spectroscopy with position-specific labeled protein substrates Reviewed

    Hidetaka Nakata, Takashi Ohtsuki, Masahiko Sisido

    Analytical Biochemistry   390 ( 2 )   121 - 125   2009.7

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    DOI: 10.1016/j.ab.2009.03.049

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  • PNA Arrays for miRNA Detection Reviewed

    Tamaki Endoh, Mizuki Kitamatsu, Masahiko Sisido, Takashi Ohtsuki

    Chemistry Letters   38 ( 5 )   438 - 439   2009.5

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    DOI: 10.1246/cl.2009.438

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  • Chemical Synthesis and Properties of 5-Taurinomethyluridine and 5-Taurinomethyl-2-thiouridine Reviewed

    Toshihiko Ogata, Tomomi Shimazaki, Tadashi Umemoto, Shinya Kurata, Takashi Ohtsuki, Tsutomu Suzuki, Takeshi Wada

    Journal of Organic Chemistry   74 ( 6 )   2585 - 2588   2009.3

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    DOI: 10.1021/jo802697r

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  • Isolation of small RNAs using biotinylated PNAs Reviewed

    Takashi Ohtsuki, Takeshi Fujimoto, Maya Kamimukai, Chisato Kumano, Mizuki Kitamatsu, Masahiko Sisido

    Journal of Biochemistry   144 ( 4 )   415 - 418   2008.10

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    DOI: 10.1093/jb/mvn107

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  • Amino acid sensing using aminoacyl-tRNA synthetase Reviewed

    Akimitsu Kugimiya, Miki Morii, Takashi Ohtsuki

    Analytical Biochemistry   378 ( 1 )   90 - 92   2008.7

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    DOI: 10.1016/j.ab.2008.03.037

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  • Modified uridines with c5-methylene substituents at the first position of the tRNA anticodon stabilize U center dot G wobble pairing during decoding Reviewed

    Shinya Kurata, Albert Weixlbaumer, Takashi Ohtsuki, Tomomi Shimazaki, Takeshi Wada, Yohei Kirino, Kazuyuki Takai, Kimitsuna Watanabe, V. Ramakrishnan, Tsutomu Suzuki

    J. Biol. Chem.   283 ( 27 )   18801 - 18811   2008.7

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    DOI: 10.1074/jbc.M800233200

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  • Cellular siRNA delivery mediated by a cell-permeant RNA-Binding protein and photoinduced RNA interference Reviewed

    Tamaki Endoh, Masahiko Sisido, Takashi Ohtsuki

    Bioconjugate Chemistry   19 ( 5 )   1017 - 1024   2008.5

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    DOI: 10.1021/bc800020n

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  • Elongation factor Tu mutants expand amino acid tolerance of protein biosynthesis system Reviewed

    Yoshio Doi, Takashi Ohtsuki, Yoshihiro Shimizu, Takuya Ueda, Masahiko Sisido

    J. Am. Chem. Soc.   129 ( 46 )   14458 - 14462   2007.11

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    DOI: 10.1021/ja075557u

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  • T-armless tRNAs and elongated elongation factor Tu Reviewed

    Takashi Ohtsuki, Yoh-ichi Watanabe

    IUBMB LIFE   59 ( 2 )   68 - 75   2007.2

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    DOI: 10.1080/15216540701218722

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  • Design of carrier tRNAs and selection of four-base codons for efficient incorporation of various nonnatural amino acids into proteins in Spodoptera frugiperda 21 (Sf21) insect cell-free translation system Reviewed

    Masumi Taki, Yasunori Tokuda, Takashi Ohtsuki, Masahiko Sisido

    J. Biosci. Bioeng.   102 ( 6 )   511 - 517   2006.12

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    DOI: 10.1263/jbb.102.511

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  • An evolutionary 'intermediate state' of mitochondrial translation systems found in Trichinella species of parasitic nematodes: co-evolution of tRNA and EF-Tu Reviewed

    Masashi Arita, Takuma Suematsu, Arihiro Osanai, Takashi Inaba, Haruo Kamiya, Kiyoshi Kita, Masahiko Sisido, Yoh-ichi Watanabe, Takashi Ohtsuki

    NUCLEIC ACIDS RESEARCH   34 ( 18 )   5291 - 5299   2006.10

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    DOI: 10.1093/nar/gkl526

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  • A protein extension to shorten RNA: elongated elongation factor-Tu recognizes the D-arm of T-armless tRNAs in nematode mitochondria Reviewed

    Masayuki Sakurai, Yoh-ichi Watanabe, Kinnitsuna Watanabe, Takashi Ohtsuki

    BIOCHEMICAL JOURNAL   399 ( 2 )   249 - 256   2006.10

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    DOI: 10.1042/BJ20060781

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  • Identification of the residues involved in the unique serine specificity of Caenorhabditis elegans mitochondrial EF-Tu2 Reviewed

    Aya Sato, Yoh-ichi Watanabe, Tsutomu Suzuki, Makoto Komiyama, Kimitsuna Watanabe, Takashi Ohtsuki

    Biochemistry   45 ( 36 )   10920 - 10927   2006.9

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    DOI: 10.1021/bi060536i

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  • Binding efficiency of elongation factor Tu to tRNAs charged with nonnatural fluorescent amino acids Reviewed

    H Nakata, T Ohtsuki, R Abe, T Hohsaka, M Sisido

    Analytical Biochemistry   348 ( 2 )   321 - 323   2006.1

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    DOI: 10.1016/j.ab.2005.08.008

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  • Expansion of protein biosynthesis system including nonnatural amino acids

    Takashi Ohtsuki, Yoshio Doi, Taishi Manabe, Masahiko Sisido

    2006 IEEE INTERNATIONAL SYMPOSIUM ON MICRO-NANOMECHATRONICS AND HUMAN SCIENCE   323 - +   2006

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  • Multiple incorporation of non-natural amino acids into a single protein using tRNAs with non-standard structures Reviewed

    T Ohtsuki, T Manabe, M Sisido

    FEBS Letters   579 ( 30 )   6769 - 6774   2005.12

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    DOI: 10.1016/j.febslet.2005.11.010

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  • Four-base codon/anticodon strategy and non-enzymatic aminoacylation for protein engineering with non-natural amino acids Reviewed

    M Sisido, K Ninomiya, T Ohtsuki, T Hohsaka

    Methods   36 ( 3 )   270 - 278   2005.7

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    DOI: 10.1016/j.ymeth.2005.04.009

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  • Isolation and physiochemical properties of protein-rich nematode mitochondrial ribosomes Reviewed

    F Zhao, T Ohtsuki, K Yamada, S Yoshinari, K Kita, Y Watanabe, K Watanabe

    Biochemistry   44 ( 25 )   9232 - 9237   2005.6

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    DOI: 10.1021/bi047833c

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  • Unusual usage of wobble modifications in mitochondrial tRNAs of the nematode Ascaris suum Reviewed

    M Sakurai, T Ohtsuki, T Suzuki, K Watanabe

    FEBS Letters   579 ( 13 )   2767 - 2772   2005.5

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    DOI: 10.1016/j.febslet.2005.04.009

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  • Modification at position 9 with 1-methyladenosine is crucial for structure and function of nematode mitochondrial tRNAs lacking the entire T-arm Reviewed

    M Sakurai, T Ohtsuki, K Watanabe

    Nucleic Acids Research   33 ( 5 )   1653 - 1661   2005

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    DOI: 10.1093/nar/gki309

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  • A unique tRNA recognition mechanism of Caenorhabditis elegans mitochondrial EF-Tu2 Reviewed

    T Suematsu, A Sato, M Sakurai, K Watanabe, T Ohtsuki

    Nucleic Acids Research   33 ( 15 )   4683 - 4691   2005

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nar/gki784

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  • Glimpses of transitions from the RNA world to the RNP world in protein compensation for RNA deficit in animal mitochondrial translation systems. Reviewed

    Suzuki, T, Ohtsuki, T, Watanabe, K

    Endocytobiosis Cell Research   2004

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  • Identification and characterization of an intermediate in the alkali degradation of (6-4) photoproduct-containing DNA Reviewed

    M Higurashi, T Ohtsuki, A Inase, R Kusumoto, C Masutani, F Hanaoka, S Iwai

    J. Biol. Chem.   278 ( 51 )   51968 - 51973   2003.12

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    DOI: 10.1074/jbc.M307186200

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  • Quick two-step RNA ligation employing periodate oxidation Reviewed

    S Kurata, T Ohtsuki, T Suzuki, K Watanabe

    Nucleic Acids Research   31 ( 22 )   2003.11

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    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/nar/gng145

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  • A unique serine-specific elongation factor Tu found in nematode mitochondria (vol 9, pg 669, 2002) Reviewed

    T Ohtsuki, A Sato, Y Watanabe, K Watanabe

    Nature Structural Biology   10 ( 8 )   669 - 669   2003.8

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    Authorship:Lead author   Language:English  

    DOI: 10.1038/nsb826

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  • Towards artificial repair of UV-damaged DNA: studies on drug binding and alkali hydrolysis. Reviewed

    S. Iwai, A. Inase, S. Jafar, M. Higurashi, T. Ohtsuki, Y. Xu, H. Sugiyama

    Nucleic acids research. Supplement (2001)   ( 3 )   181 - 182   2003

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  • Decoding property of C5 uridine modification at the wobble position of tRNA anticodon. Reviewed

    Kurata S, Ohtsuki T, Wada T, Kirino Y, Takai K, Saigo K, Watanabe K, Suzuki T

    Nucleic acids research. Supplement (2001)   ( 3 )   245 - 246   2003

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  • Characterization of the interaction between the nucleotide exchange factor EF-Ts from nematode mitochondria and elongation factor Tu Reviewed

    T Ohtsuki, M Sakurai, A Sato, K Watanabe

    Nucleic Acids Research   30 ( 24 )   5444 - 5451   2002.12

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    DOI: 10.1093/nar/gkf679

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  • Solution structure of an RNA fragment with the P7/P9.0 region and the 3 '-terminal guanosine of the Tetrahymena group I intron Reviewed

    A Kitamura, Y Muto, S Watanabe, Kim, I, T Ito, Y Nishiya, K Sakamoto, T Ohtsuki, G Kawai, K Watanabe, K Hosono, H Takaku, E Katoh, T Yamazaki, T Inoue, S Yokoyama

    RNA   8 ( 4 )   440 - 451   2002.4

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    DOI: 10.1017/S1355838202026043

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  • The minimal tRNA: unique structure of Ascaris suum mitochondrial tRNA(UCU)(Ser) having a short T arm and lacking the entire D arm Reviewed

    T Ohtsuki, G Kawai, K Watanabe

    FEBS Letters   514 ( 1 )   37 - 43   2002.3

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  • An unnatural base pair for incorporating amino acid analogs into proteins Reviewed

    Hirao, I, T Ohtsuki, T Fujiwara, T Mitsui, T Yokogawa, T Okuni, H Nakayama, K Takio, T Yabuki, T Kigawa, K Kodama, T Yokogawa, K Nishikawa, S Yokoyama

    Nature Biotechnology   20 ( 2 )   177 - 182   2002.2

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    DOI: 10.1038/nbt0202-177

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  • An "elongated" translation elongation factor Tu for truncated tRNAs in nematode mitochondria Reviewed

    T Ohtsuki, Y Watanabe, C Takemoto, G Kawai, T Ueda, K Kita, S Kojima, Y Kaziro, J Nyborg, K Watanabe

    J. Biol. Chem.   276 ( 24 )   21571 - 21577   2001.6

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    DOI: 10.1074/jbc.M011118200

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  • Unnatural base pairs for specific transcription Reviewed

    T Ohtsuki, M Kimoto, M Ishikawa, T Mitsui, Hirao, I, S Yokoyama

    Proc. Natl. Acad. Sci. U.S.A.   98 ( 9 )   4922 - 4925   2001.4

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    DOI: 10.1073/pnas.091532698

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  • Dual specificity of the pyrimidine analogue, 4-methylpyridin-2-one, in DNA replication Reviewed

    Hirao, I, T Ohtsuki, T Mitsui, S Yokoyama

    J. Am. Chem. Soc.   122 ( 25 )   6118 - 6119   2000.6

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  • The guanosine binding mechanism of the Tetrahymena group I intron

    A. Kitamura, Y. Muto, S. Watanabe, I. Kim, T. Ito, Y. Nishiya, T. Ohtsuki, G. Kawai, K. Watanabe, K. Hosono, H. Takaku, E. Katoh, T. Yamazaki, T. Inoue, S. Yokoyama

    Nucleic Acids Symposium Series   42 ( 1 )   191 - 192   1999.11

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    Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

    DOI: 10.1093/nass/42.1.191

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  • Stable isotope edited NMR analysis of Ascaris suum mitochondrial tRNA(Met) having a TV-replacement loop Reviewed

    T Ohtsuki, G Kawai, K Watanabe

    Journal of Biochemistry   124 ( 1 )   28 - 34   1998.7

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  • Automated chemical synthesis of biologically active tRNA having a sequence corresponding to Ascaris suum mitochondrial tRNA(Met) toward NMR measurements Reviewed

    T Ohtsuki, R Vinayak, Y Watanabe, K Kita, G Kawai, K Watanabe

    Journal of Biochemistry   120 ( 6 )   1070 - 1073   1996.12

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  • Preparation of biologically active Ascaris suum mitochondrial tRNA(Met) with a TV-replacement loop by ligation of chemically synthesized RNA fragments Reviewed

    T Ohtsuki, G Kawai, Y Watanabe, K Kita, K Nishikawa, K Watanabe

    Nucleic Acids Research   24 ( 4 )   662 - 667   1996.2

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  • Mollification of Cytotoxicity of Sulfated Polysaccharides by Fibroblast Growth Factors Reviewed

    M KUNOU, T OHTSUKI, T AKAIKE, K HATANAKA

    Journal of Carbohydrate Chemistry   14 ( 4-5 )   659 - 665   1995

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  • Effects of Synthetic Polyanions on 3T3-L1 Fibroblast Proliferation Stimulated by Fibroblast Growth Factors Reviewed

    K HATANAKA, T OHTSUKI, M KUNOU

    Chemistry Letters   1407-1410 ( 8 )   1407 - 1410   1994.8

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    DOI: 10.1246/cl.1994.1407

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▼display all

Books

  • 化学の指針シリーズ「生物有機化学 -ケミカルバイオロジーへの展開-」

    宍戸昌彦,大槻高史

    裳華房  2008 

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  • 新規モダリティ医薬品のための新しいDDS技術と製剤化

    著者多数(含:大槻高史)( Role: Joint author ,  高分子ナノミセル・ペプチド-RNA ナノ複合体を基盤としたキャリア作製、ペプチドジッパー法による細胞膜透過ペプチドを用いたカーゴの細胞内デリバリー)

    技術情報協会  2023.1  ( ISBN:9784861049361

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    Total pages:580p   Language:Japanese

    CiNii Books

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  • 核酸科学ハンドブック(共著)

    著者多数(含:大槻高史)( Role: Joint author ,  RNAデリバリー)

    講談社サイエンティフク  2020.12 

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  • CSJ current review: 生体分子反応を制御する(共著)

    著者多数(含:大槻高史)( Role: Joint author ,  非天然アミノ酸による生体反応の制御法)

    化学同人  2020.4 

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  • ペプチド創薬の最前線 = The frontier of peptide drug discovery

    木曽, 良明( Role: Joint author ,  ペプチドファスナー法による生理活性ペプチドの細胞内デリバリー)

    シーエムシー出版  2019.5  ( ISBN:9784781314174

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    Total pages:viii, 265p   Language:Japanese

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  • ペプチド医薬品のスクリーニング・安定化・製剤化技術(共著)

    技術情報協会  2017 

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  • Nanotechnology Tools for the Study of RNA(共著)

    Elsevier  2014 

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  • Intracellular Delivery II

    Kazunori Watanabe, Takashi Ohtsuki( Role: Joint author ,  Intracellular delivery of RNA via RNA-binding proteins or peptides. pp 403-416)

    2014  ( ISBN:9789401788953

  • 人工核酸によるRNAの検出と精製,In 「シングルセル解析の最前線」

    シーエムシー出版  2010 

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  • RNA Interference

    Springer  2010 

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  • The central dogma: from DNA to RNA, and to protein. In “Automation in Proteomics and Genomics”

    ( Role: Joint author)

    Wiley  2009 

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  • 蛋白質核酸酵素増刊「RNAの細胞生物学」Vol.48, No.4

    共立出版  2003 

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  • 日本臨床増刊「ミトコンドリアとミトコンドリア病」,Vol. 60, No.4

    日本臨床社  2002 

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  • Cell-free translation system

    Springer-Verlag  2002 

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  • 基礎生化学実験法 第4巻

    東京化学同人  2000 

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MISC

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Presentations

  • Cell-penetrating peptide/photosensitizer conjugates for photo-triggered cytosolic delivery of RNAs Invited

    Takashi Ohtsuki

    Asia 3 Round Table on Nucleic Acids  2024.11.3 

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    Event date: 2024.11.1 - 2024.11.3

    Language:English   Presentation type:Oral presentation (invited, special)  

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  • Cell-penetrating peptide/photosensitizer conjugates for photo-triggered cytosolic delivery of RNAs and peptides Invited

    Takashi Ohtsuki

    18th International Congress on Photobiology  2024.8.27 

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    Event date: 2024.8.26 - 2024.8.30

    Language:English   Presentation type:Oral presentation (invited, special)  

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  • RNA encapsulation and photo-responsive release by caged coacervates based on liquid-liquid phase separation

    坂東晃成, 渡邉和則, 金崎佑紀, 北松瑞生, 大槻高史

    日本化学会春季年会講演予稿集(Web)  2024 

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  • Development of the analyzing method for single nucleotide-mutated single-cancer cells using Peptide Nucleic acid Probes and Single-Cell Microarray

    重藤元, 北松瑞生, 大槻高史, 飯塚明, 秋山靖人, 山村昌平

    日本化学会春季年会講演予稿集(Web)  2024 

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  • Development of biodegradable micellar boron preparation for BNCT Therapy

    井上晴喜, FITHRONI Abdul Basith, 渡邉和則, 松浦栄次, 大槻高史

    日本バイオマテリアル学会大会予稿集(Web)  2023 

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  • Development of photothermal therapy agents that added tumor specificity and improved apoptosis induction efficiency

    杉原桃香, 大槻高史, 渡邉和則

    日本バイオマテリアル学会大会予稿集(Web)  2023 

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  • 哺乳動物初期胚への光による時空間特異的なRNA導入法

    大槻高史, 井川優風, 若井拓哉, 舟橋弘晃, 渡邉和則

    日本光医学・光生物学会  2023 

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  • 光化学的内在化法の副作用の低減

    坂東晃成, 渡邉和則, 大槻高史

    日本化学会春季年会講演予稿集(Web)  2023 

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  • 光増感剤と核酸キャリアの併用による光依存的mRNAデリバリー法

    前本隼玖, 渡邉和則, 位高啓史, 大槻高史

    日本光医学・光生物学会  2023 

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  • Elucidation of Ca2+ signaling mechanisms in heat stress-dependent nuclear stress bodies formation

    古谷優治, 的野恭平, 大槻高史, 渡邉和則

    日本分子生物学会年会プログラム・要旨集(Web)  2023 

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  • Elucidation of relationship between thermo-tolerance and cell death via translational suppression and initiator tRNA degradation

    青木結子, 梅本裕介, 長房すずか, 高橋昭久, 井尻憲一, 大槻高史, 渡邉和則

    日本分子生物学会年会プログラム・要旨集(Web)  2023 

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  • Elucidation of Xrn1 and Xrn2 activation mechanism related to initiator tRNA degradation by heat stress

    渡邉和則, 岩城香菜子, 篠原里菜, 安田奈緒, 甲斐健太郎, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2023 

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  • 熱ストレス下でのHSF1リン酸化が顆粒形成に及ぼす影響

    小笠原悠人, 大槻高史, 渡邉和則

    日本臨床ストレス応答学会大会抄録集  2023 

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  • Reduction of side effects of the PCI method

    坂東晃成, 渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2022 

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  • Spatiotemporally photocontrolled RNA transfection in early mammalian embryos

    井川優風, 若井拓哉, 舟橋弘晃, 渡邉和則, 大槻高史

    日本バイオマテリアル学会大会予稿集(Web)  2022 

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  • Biodegradable nanocarriers for efficient light-dependent intracellular delivery of siRNA.

    田中七星, 渡邉和則, 小渕浩嗣, 久保貴紀, 松浦栄次, 大槻高史

    日本バイオマテリアル学会大会予稿集(Web)  2022 

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  • Relationship between tRNA Degradation by heat stress and Cell Cycle-Dependent Translational suppression

    竹本理恵, 梅本裕介, 長房すずか, 高橋昭久, 井尻憲一, 大槻高史, 渡邉和則

    日本分子生物学会年会プログラム・要旨集(Web)  2022 

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  • Inhibition of HSF1 and SAFB granule formation enhances apoptosis by heat stress

    渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2022 

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  • RNA分解を検出する蛍光寿命プローブ

    平田 陸, 平川 和貴, 島田 尚鷹, 渡邉 和則, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 相分離生物学 相分離を介した核内ストレス顆粒の形成は温熱抵抗性に関与している

    渡邉 和則, 的野 恭平, 大槻 高史

    Thermal Medicine  2021.9  (一社)日本ハイパーサーミア学会

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  • Photocontrolled apoptosis induction using pre-miR-664a and PCDR method

    渡邉和則, 渡邉和則, 縄稚朋子, 奥谷瑠璃子, 大槻高史, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2021 

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  • Evaluation of secondary structure and intracellular delivery by stapling of apoptosis-inducing peptide Bim

    西村風香, 北松瑞生, 藤井香帆, 大槻高史

    日本化学会春季年会講演予稿集(Web)  2021 

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  • mTOR complex1 was involved in the formation of SAFB granules under heat stress

    的野恭平, 井上歩実, 岡田真実, 山本理紗子, 大槻高史, 渡邉和則

    日本分子生物学会年会プログラム・要旨集(Web)  2021 

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  • 細胞内へのタンパク質輸送のためPCI分子素子の開発

    角菜々子, 北松瑞生, 阿部由佳, 渡邉和則, 大槻高史

    日本生化学会大会(Web)  2021 

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  • RNA分解を検出する蛍光寿命プロープ

    平田陸, 平川和貴, 島田尚鷹, 渡邉和則, 大槻高史

    日本生化学会大会(Web)  2021 

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  • miRNA detection by Fluorophore-PNA-Quencher/Quencher-DNA probes

    田原健太朗, 渡邉和則, 重藤元, 山村昌平, 岸高稚, 北松瑞生, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2021 

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  • 核内ストレス顆粒の形成は温熱抵抗性に関与している

    渡邉和則, 大槻高史

    Thermal Medicine  2020 

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  • mTOR複合体を介した核内ストレス顆粒構成因子SAFB,Satellite III RNA顆粒形成機構の解明

    的野恭平, 井上歩実, 岡田真実, 山本理紗子, 大槻高史, 大槻高史, 渡邉和則, 渡邉和則

    Thermal Medicine  2020 

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  • 温熱によるXrn1/2活性化機構の解明

    岩城 香菜子, 大槻 高史, 渡邉 和則

    Thermal Medicine  2019.9  (一社)日本ハイパーサーミア学会

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  • 温熱によるSatellite III RNAの発現上昇はmTOR複合体により制御されている

    渡邉和則, 井上歩実, 大槻高史

    日本RNA学会年会要旨集  2019 

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  • 光応答的に働く卵活性化因子の開発

    大本和正, 若井拓哉, 舟橋弘晃, 渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2019 

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  • mTOR複合体制御による核内ストレス顆粒形成機構の解明

    渡邉和則, 井上歩実, 岡田真実, 山本理紗子, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2019 

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  • mTOR複合体を介した核内ストレス顆粒形成機構の解明

    渡邉和則, 井上歩実, 岡田真実, 山本理紗子, 大槻高史

    Thermal Medicine  2019 

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  • 光応答性キャリアを用いたphotochemical internalizationの機構解明

    南條 友孝, 渡邉 和則, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2018.9  (公社)日本生化学会

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  • 光に応答し細胞質内に侵入する機能性光増感剤ペプチドの設計

    三好 祐一, 山本 怜見, 門野 真保, 北松 瑞生, 渡邉 和則, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2018.9  (公社)日本生化学会

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  • 光依存的なRNA導入法による2種類のRNAの時間差導入

    大槻 高史, 白神 かおり, 渡邉 和則

    日本生化学会大会プログラム・講演要旨集  2018.9  (公社)日本生化学会

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  • 細胞内のRNA分解に関する蛍光相関分析法による解析(Fluorescence correlation spectroscopy analysis of RNA degradation in cells)

    島田 尚鷹, 渡邉 和則, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2018.9  (公社)日本生化学会

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  • mTOR複合体を介した温熱による核内ストレス構造体形成機構の解明

    井上 歩実, 岡田 真実, 山本 理紗子, 大槻 高史, 渡邉 和則

    Thermal Medicine  2018.8  (一社)日本ハイパーサーミア学会

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    Language:Japanese  

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  • 温熱による開始tRNA分解を介した翻訳抑制と細胞周期との関係性

    渡邉 和則, 梅本 裕介, 長房 すずか, 高橋 昭久, 井尻 憲一, 大槻 高史

    Thermal Medicine  2018.8  (一社)日本ハイパーサーミア学会

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  • 熱ストレスによる核内ストレス構造体の形成はmTOR複合体によって制御されている

    井上歩実, 岡田真実, 山本理紗子, 大槻高史, 渡邉和則

    日本生化学会大会(Web)  2018 

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  • 温熱により形成される核内ストレス顆粒形成機構の解明

    渡邉和則, 井上歩実, 岡田真実, 山本理紗子, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2018 

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  • 細胞周期依存的な開始tRNA分解を介した温熱による翻訳抑制機構

    渡邉和則, 渡邉和則, 梅本裕介, 長房すずか, 高橋昭久, 井尻憲一, 大槻高史

    日本生化学会大会(Web)  2018 

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  • Pre-miR-664aを用いた光依存的な細胞死の誘導

    縄稚 朋子, 渡邉 和則, 大槻 高史

    生命科学系学会合同年次大会  2017.12  生命科学系学会合同年次大会運営事務局

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    Language:Japanese  

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  • mTORを介した核内ストレス構造体の熱ストレス下における形成機構

    井上 歩実, 渡邉 和則, 岡田 真実, 山本 理紗子, 大槻 高史

    生命科学系学会合同年次大会  2017.12  生命科学系学会合同年次大会運営事務局

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    Language:Japanese  

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  • mTORを介した温熱による核内ストレス構造体形成機構

    渡邉 和則, 岡田 真実, 井上 歩実, 山本 理紗子, 大槻 高史

    Thermal Medicine  2017.9  (一社)日本ハイパーサーミア学会

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  • 音増感剤を用いた超音波依存的なペプチドの細胞質内導入法

    大槻 高史, 稲葉 優樹, 北松 瑞生, 中田 栄司, 原田 敦史, 渡邉 和則

    日本DDS学会学術集会プログラム予稿集  2017.6  日本DDS学会

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    Language:Japanese  

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  • ペプチドとRNA双方でアポトーシスを誘導するナノ複合体の開発

    金 亨振, 北松 瑞生, 大槻 高史

    日本DDS学会学術集会プログラム予稿集  2017.6  日本DDS学会

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  • P20 Ultrasound-dependent cytosolic delivery of bioactive molecules

    Nagahiro Shota, Inaba Yuki, Matuura Eiji, Watanabe Kazunori, Ohtsuki Takashi

    Proceedings of the Annual Meeting of the Japan Society of Sonochemistry  2017  Japan Society of Sonochemistry

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    Recently, methods for delivery of peptide and RNA to target cells are widely studied for cell research and drug delivery strategies. However, the delivery methods sometimes have problems such as cytotoxicity and ineffective delivery. In some methods, target molecules tend to be trapped in endosome and degraded by lysosome when introduced to cells. In this study, ultrasound-dependent peptide / nanoparticle delivery method was developed. We developed a molecular conjugate of a cell-penetrating peptide, a sonosensitizer and a functional peptide / nanoparticle. This molecule was trapped in endosome when cells were incubated with the molecule. However, we succeeded in cytoplasmic delivery of the molecule that escape from endosome by ultrasound irradiation.

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  • 高分子ミセル「ラクトソーム」を用いた光照射特異的なRNAi技術の開発

    大島 真, 赤星 彰也, 松浦 栄次, 小関 英一, 渡邉 和則, 大槻 高史

    日本バイオマテリアル学会大会予稿集  2016.11  日本バイオマテリアル学会

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    Language:Japanese  

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  • 熱ストレスによる核内ストレス構造体形成機構の解明

    渡邉 和則, 岡田 真実, 山本 理紗子, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2016.9  (公社)日本生化学会

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    Event date: 2016.9

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  • 熱ストレス依存的な核内ストレス構造体形成機構の解析

    岡田真実, 山本理沙子, 渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2016 

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  • 細胞機能を解析し創る新技術 1分子から時間空間制御まで 可視光および近赤外光による細胞内RNA導入の局所的誘導

    大槻 高史, 白神 かおり, 渡邉 和則

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  2015.12  (公社)日本生化学会

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  • RNAの分解と寿命の解明

    大西 諒, 中林 孝和, 渡邉 和則, 大槻 高史

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  2015.12  (公社)日本生化学会

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  • 新規神経分化関連microRNAの探索

    山路 隆平, 渡邉 和則, 大槻 高史

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  2015.12  (公社)日本生化学会

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  • 光でアポトーシスを誘導する方法の開発

    藤原隼人, 畑地祐里, 北松瑞生, 渡邉和則, 大槻高史

    日本化学会中国四国支部大会講演要旨集  2015.11.7 

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  • 拡張翻訳系を用いた糖タンパク質作製法 Invited

    大槻 高史, 矢形 梓, 白神 かおり, 渡邉 和則, 畑中 研一

    第34回日本糖質学会年会  2015.7.31 

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  • 熱ストレスによる開始tRNAMetは細胞周期に依存して分解促進している

    渡邉和則, 渡邉和則, 梅本裕介, 高橋昭久, 井尻憲一, 大槻高史

    日本RNA学会年会要旨集  2015 

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  • ケージドアミノアシルtRNAを用いた翻訳の光制御法の開発

    神崎重人, 渡邉和則, 大槻高史

    日本化学会中国四国支部大会講演要旨集  2015 

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  • 光増感剤と高分子ミセルを用いたRNAデリバリーキャリアの設計

    井野川 翔一, 渡邊 和則, 小関 英一, 松浦 栄次, 大槻 高史

    日本バイオマテリアル学会大会予稿集  2014.11  日本バイオマテリアル学会

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    Language:Japanese  

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  • 光化学的に細胞質内に侵入するペプチド分子の設計

    大槻高史, 藤原隼人, 畑地祐里, 北松瑞生, 渡邉和則

    光化学討論会要旨集(CD-ROM)  2014 

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  • tRNA顆粒形成要因タンパク質同定の試み

    山本理紗子, 大槻高史, 渡邉和則

    日本分子生物学会年会プログラム・要旨集(Web)  2014 

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  • mTORが熱ストレスによる開始tRNAMetの分解促進を制御している

    渡邉和則, 渡邉和則, 井尻憲一, 大槻高史

    日本RNA学会年会要旨集  2014 

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  • 熱ストレス下では,mTORはXrn2の核小体から核質への拡散を介して開始tRNAMetの分解促進を制御している

    渡邉和則, 渡邉和則, 井尻憲一, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2014 

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  • CLIP-RNAi法における多環状RNAの有効性の検討

    村上真一, 渡邉和則, 大槻高史

    日本RNA学会年会要旨集  2014 

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  • 細胞内運搬ペプチドと光応答色素を含むアポトーシス誘導ペプチドの合成とそのエンドソーム内蓄積

    前場伊織, 北松瑞生, 大槻高史

    日本化学会講演予稿集  2013.3.8 

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  • ヘテロ二量体化ロイシンジッパーによる緑色蛍光タンパク質の細胞内輸送

    出口宝, 北松瑞生, 中島真実, 大槻高史, 道上宏之

    日本化学会講演予稿集  2013.3.8 

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  • 細胞周期とRNA関連イベントとの相関の観察手法の開発

    丸田静佳, 渡邉和則, 大槻高史

    アンチセンスシンポジウム講演要旨集  2013 

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  • 多環状RNAを用いた光誘導RNAi

    村上真一, 渡邉和則, 大槻高史

    日本分子生物学会年会プログラム・要旨集(Web)  2013 

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  • ケージドアミノアシルtRNAを用いた翻訳の光制御

    赤星彰也, 土井芳朗, 木内智樹, 渡邉和則, 大槻高史

    日本RNA学会年会要旨集  2013 

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  • 細胞内noncoding RNA局在追跡ツールの開発

    田村 佳史, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2012.12  (公社)日本生化学会

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    Language:Japanese  

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  • 分岐状アミノ酸と蛍光性アミノ酸から成るペプチドデンドリマーの合成およびその性質

    北松瑞生, 北畠まゆみ, 能年義輝, 大槻高史

    高分子学会予稿集(CD-ROM)  2012.9.5 

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    Event date: 2012.9.5

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  • 中心に蛍光基を含んだ分岐状ペプチドの性質

    北畠まゆ美, 脇坂祐也, 石踊由佳, 大槻高史, 北松瑞生

    日本化学会西日本大会講演要旨集  2011.11.7 

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  • 細胞内侵入型PNAビーコンによる機能性RNAの検出

    西岡浩貴, 大槻高史, 北松瑞生

    日本化学会西日本大会講演要旨集  2011.11.7 

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  • 細胞内侵入型PNAによる機能性RNAの導入および検出

    西岡浩貴, 大槻高史, 北松瑞生

    アンチセンスシンポジウム講演要旨集  2011.9.1 

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  • アルギニン誘導体の部位特異的導入に基づくヒストン蛋白質の機能解析

    赤星 彰也, 鈴江 良隆, 宍戸 昌彦, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2011.9  (公社)日本生化学会

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  • リポソーム膜上における膜蛋白質への蛍光性アミノ酸導入およびその挙動解析法の開発

    大塚 巧磨, 根木 慧, 金井 保, 秋吉 一成, 野村 M. 新一郎, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2011.9  (公社)日本生化学会

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  • 近赤外光による照射領域特異的RNAi誘導法の開発

    石躍 由佳, 森長 美香, 大槻 高史

    日本生化学会大会プログラム・講演要旨集  2011.9  (公社)日本生化学会

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  • Synthesis of cyclic peptides using the NEXT-A cyclization reaction

    HAMAMOTO Toshimasa, SISIDO Masahiko, OHTSUKI Takashi, TAKI Masumi

    2011.3.1 

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  • CLIP-RNAi法によるRNAiの光誘導

    大槻高史, 米澤政樹, 松本祥, 遠藤玉樹, 石躍由佳

    日本光医学・光生物学会  2011 

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  • ケージドアミノアシルtRNAを用いた翻訳の光制御

    土井芳朗, 赤星彰也, 宍戸昌彦, 大槻高史

    日本化学会バイオテクノロジー部会シンポジウム講演要旨集  2011 

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  • 光応答性キャリア蛋白質を用いたRNAデリバリー

    大槻高史, 松本祥, 遠藤玉樹, 石躍由佳

    アンチセンスシンポジウム講演要旨集  2011 

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  • 細胞質内RNA導入の光誘導とCLIP-RNAi

    大槻高史, 石躍由佳, 遠藤玉樹, 宍戸昌彦

    日本RNA学会年会要旨集  2010 

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  • 光照射領域特異的RNAi誘導技術の開発

    石躍由佳, 桑原里奈, 米澤政樹, 遠藤玉樹, 大槻高史

    アンチセンスシンポジウム講演要旨集  2010 

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  • CLIP-RNAi法による遺伝子発現抑制の光誘導

    大槻高史, 石躍由佳, 遠藤玉樹, 宍戸昌彦

    日本化学会バイオテクノロジー部会シンポジウム講演要旨集  2010 

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  • CLIP-RNAi法による光照射領域特異的な遺伝子発現抑制

    大槻 高史, 遠藤 玉樹, 宍戸 昌彦

    日本生化学会大会プログラム・講演要旨集  2009.9  (公社)日本生化学会

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  • CLIP-RNAi法による1細胞RNAiの経時的観察

    大槻 高史, 遠藤 玉樹, 宍戸 昌彦

    バイオイメージング  2009.7.15 

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  • TatEF-Tuを用いたアミノアシルtRNAの哺乳動物細胞内への輸送

    根木慧史, 大槻高史, 宍戸昌彦

    日本分子生物学会年会講演要旨集  2009 

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  • 光操作による哺乳細胞内での蛋白質発現制御

    土井芳朗, 大槻高史, 宍戸昌彦

    日本分子生物学会年会講演要旨集  2009 

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  • siRNAデリバリーのためのCPP-RBP型RNAキャリアの開発

    桑原里奈, 遠藤玉樹, 米澤政樹, 宍戸昌彦, 大槻高史

    日本分子生物学会年会講演要旨集  2009 

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  • ペプチド核酸‐細胞内導入ペプチドコンジュゲートを用いたRNAの細胞内送達

    久保貴紀, 北松瑞生, 遠藤玉樹, 山中智史, 大庭秀樹, 大槻高史, 宍戸昌彦

    日本化学会西日本大会講演要旨集  2007.11.10 

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  • ペプチド核酸‐膜透過ペプチド オリゴマーをキャリアーとして用いたsiRNAの細胞内輸送

    北松瑞生, 久保貴紀, 大庭英樹, 遠藤玉樹, 大槻高史, 宍戸昌彦

    バイオ・高分子シンポジウム講演要旨集  2007.7.23 

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    Event date: 2007.7.23

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  • Antisense Effects of Pyrrolidine-Based Oxy-PNAs Conjugated with a Membrane-Permeable Peptide in Escherichia coli

    KURAMI Syunsuke, KITAMATSU Mizuki, OHTSUKI Takashi, SISIDO Masahiko

    2007.3.1 

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  • ペプチド核酸-細胞内導入ペプチドコンジュゲートを用いたRNAの細胞内送達

    久保貴紀, 北松瑞生, 遠藤玉樹, 山中智史, 大庭秀樹, 大槻高史, 宍戸昌彦

    日本化学会西日本大会講演要旨集  2007 

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  • Specific RNA binding of immobilized PNA's

    T. Ohtsuki, T. Fujimoto, Y. Sugimoto, M. Kitamatsu, M. Sisido

    JOURNAL OF PEPTIDE SCIENCE  2006  JOHN WILEY & SONS LTD

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  • interactions between nematode mitochondrial tRNAs and EF-Tus

    OHTSUKI Takashi, WATANABE Yoh-ichi, KITA Kiyoshi, TAKEMOTO Chie, KAWAI Gota, UEDA Takuya, WATANABE Kimituna

    1998.12.1 

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    Language:Japanese  

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  • NMR analysis of guanosine binding mechanism of the group I intoron

    KITAMURA Aya, MUTO Yutaka, KIM Insil, WATANABE Satoru, OOTSUKI Takashi, HOSONO Kazumi, KAWAI Gota, WATANABE Kimitsuna, TAKAKU Hiroshi, INOUE Tan, YOKOYAMA Shigeyuki

    1998.12.1 

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    Language:Japanese  

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  • 安定同位体標識NMR法による線形動物ミトコンドリアtRNAの立体構造解析

    大槻 高史, 河合 剛太, 渡辺 洋一, 西川 一八, 渡辺 公綱

    日本分子生物学会年会プログラム・講演要旨集  1996.8.1 

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    Language:Japanese  

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  • 光化学的内在化によるRNAやペプチド/タンパク質の細胞質内送達 Invited

    大槻 高史

    第46回日本光医学・光生物学会  2024.7.6 

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  • 光で細胞機能を操るための光応答分子の開発 Invited

    大槻高史

    第13回徳島文理大学 薬学部学術講演会  2020.2.5 

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  • 光で細胞機能を操るための光応答分子の開発 Invited

    大槻高史

    第24回関西大学先端科学技術シンポジウム  2020.1.23 

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  • Photochemical and sonochemical internalization of CPP-cargo-sensitizer conjugates Invited

    T. Ohtsuki

    Pacific Rim Nano Medicine Symposium 2018 -The 9th Japan-Taiwan Symposium on Nanomedicine  2018.1.25 

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  • Photochemical internalization of cell penetrating peptide-cargo-photosensitizer conjugates Invited

    Takashi Ohtsuki

    10th International Symposium on Nanomedicine  2016.11.25 

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  • 可視光および近赤外光による細胞内RNA導入の局所的誘導 Invited

    大槻高史, 白神かおり, 渡邉和則

    BMB2015(分子生物学会・生化学会)  2015.12.2 

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  • 光を用いた細胞内への物質導入 Invited

    大槻 高史

    新学術領域研究「ナノメディシン分子科学」+「超高速バイオアセンブラ」合同若手の会  2015.10.31 

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  • 光に応答する生体機能分子 Invited

    大槻高史

    第2回 日本バイオマテリアル学会中四国シンポジウム  2014.2.28 

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  • 光で細胞機能を操る Invited

    大槻高史

    テラヘルツと生命科学融合による革新的イノベーションワークショップ  2013.12.5 

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  • Light-directed RNAi using a photosensitive carrier molecule Invited

    Takashi Ohtsuki

    International Symposium on Nanomedicine Molecular Science  2013.10.8 

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  • RNAの細胞内導入と RNA機能の光誘導 Invited

    大槻高史

    生化学若い研究者の会 中四国支部 夏のセミナー  2011.8.6 

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  • Spatial regulation of specific gene expression through photoactivation of RNAi. Invited

    Takashi Ohtsuki

    2010.9.9 

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  • アミノアシルtRNA合成酵素によるアミノ酸認識特異性 Invited

    大槻高史

    バイオ分析研究会  2009.12.11 

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  • Applications of peptide nucleic acids for RNA purification, RNA detection, and intracellular RNA delivery Invited

    Takashi Ohtsuki

    The 6'th International Forum On Post-Genome Technologies  2009.9.16 

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  • RNAi創薬を目指した新規RNAキャリアの開発 Invited

    大槻高史

    第2回高度医療都市を創出する未来技術国際シンポジウム  2009.2.5 

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  • Position specific incorporation of nonnatural amino acids into proteins in vivo

    JAACT2008  2008 

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  • tRNA構造との対応関係を越えたショウジョウバエミトコンドリアEF-TuのtRNA認識能

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • Improvement of CPP-RBD for siRNA delivery

    JAACT2008  2008 

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  • アルギニン誘導体の蛋白質への位置特異的導入

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • 哺乳細胞を用いた非天然アミノ酸導入蛋白質合成法の開発

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • RNA-蛋白質ハイブリッド型FRETプローブを用いた低分子化合物の検出

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • EF-Tu変異体によるアミノアシルtRNAの精製・定量法

    第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会  2008 

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  • EF-Tu変異体を用いた非天然アミノアシルtRNAの精製と定量

    第3回バイオ関連化学合同シンポジウム  2008 

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  • 細胞内での非天然アミノ酸導入蛋白質合成法の開発」第3回バイオ関連化学合同シンポジウム

    第3回バイオ関連化学合同シンポジウム  2008 

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  • Construction of RNA-protein Hybrid for Bioimaging of Low-molecular Compounds.

    3rd International Workshop on Approaches to Single-Cell Analysis  2008 

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  • CLIP-RNAi ~photoinduced RNAi strategy~.

    3rd International Workshop on Approaches to Single-Cell Analysis  2008 

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  • 蛍光標識CPP-RBPによるshRNAの細胞内導入と遺伝子発現の光制御

    日本化学会第88春季年会  2008 

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  • RNA-蛋白質ハイブリッド型FRETプローブによる低分子化合物の検出

    日本化学会第88春季年会  2008 

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  • 蛍光標識CPP-RBPによるRNAiの光誘導

    第10回RNAミーティング  2008 

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  • 細胞内生理活性分子検出のためのRNA-タンパク質ハイブリッド型FRETプローブの構築

    第10回RNAミーティング  2008 

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  • 酵素反応を用いるアミノ酸の蛍光分析

    日本化学会第88春季年会  2008 

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  • 蛍光標識CPP-RBPによるRNAデリバリーとRNAiの光制御

    遺伝子・デリバリー研究会 第8回シンポジウム  2008 

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  • 蛋白質キャリアによるsiRNAの細胞内導入とRNAiの光誘導 Invited

    大槻高史

    第212回バイオロンジル会  2007.10.30 

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  • 非天然アミノ酸含有タンパク質の無細胞及び細胞モデル系での発現 Invited

    大槻高史

    特定領域『バイオ操作』若手研究者第2回ワークショップ  2007.7.20 

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  • RNA工学と拡張翻訳系 Invited

    大槻高史

    第109回応用化学科セミナー(愛媛大学)  2007.7.2 

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  • 改変EF-Tuによる蛋白質への非天然アミノ酸高効率導入

    第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会  2007 

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  • 生細胞内における蛋白質への位置特異的な蛍光基導入法の開発

    第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会  2007 

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  • アセチル化アミノ酸やメチル化アミノ酸の蛋白質への位置特異的導入

    第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会  2007 

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  • RNA interference using cell permeable protein carrier

    4th International Peptide Symposium  2007 

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  • Delivery of dsRNA with lactic acid bacteria for RNA interference

    Kuwahara, A, Arita, M, Sisido, M, Ohtsuki, T

    5th International Symposium on Nucleic Acids Chemistry  2007 

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  • Photo inducible RNA interference using cell permeable protein carrier

    Endoh, T, Sisido, M, Ohtsuki, T

    5th International Symposium on Nucleic Acids Chemistry  2007 

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  • Incorporation of large nonnatural amino acids into protein by translation with EF-Tu mutants

    4th International Peptide Symposium  2007 

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  • ISFET電極を用いるアミノ酸センサーの作製と応答の評価

    日本化学会第87春季年会  2007 

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  • RNA isolation using biotinylated PNAs

    The 2nd International Workshop on Approaches to Single Cell Analysis  2007 

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  • リボソームを用いたβ-ペプチド合成に向けたアプローチ:主鎖伸長型基質導入に関与するファクター

    第22回生体機能関連化学シンポジウム  2007 

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  • ペプチド核酸-膜透過ペプチド オリゴマーをキャリアーとして用いたsiRNAの細胞内輸送

    第17回バイオ・高分子シンポジウム  2007 

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  • 蛍光標識Tat融合RNA結合タンパク質を用いたsiRNAの細胞内導入とRNAiの光制御

    第22回生体機能関連化学シンポジウム  2007 

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  • 改変EF-Tuを用いた翻訳系の拡張

    第22回生体機能関連化学シンポジウム  2007 

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  • Cellular delivery and photo-accelerated endosomal escape of siRNA mediated by cell permeable RNA binding protein

    12th annual meeting of RNA society  2007 

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  • 蛋白質生合成の非天然アミノ酸許容性の拡張を目的とする変異EF-Tuの作製

    日本ケミカルバイオロジー第2回年会  2007 

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  • Expansion of protein biosynthesis system by EF-Tu mutants.

    12th annual meeting of RNA society  2007 

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  • Expansion of protein biosynthesis system that includes nonnatural amino acids Invited

    Takashi Ohtsuki, Yoshio Doi, Taishi Manabe, Masahiko Sisido

    Okayama Peptide Symposium 2006  2006.11.9 

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  • 無細胞蛋白質合成システムを用いた非天然アミノ酸導入蛋白質の合成とその応用 Invited

    大槻高史

    第一回無細胞生命科学研究会  2006.10.7 

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  • Expansion of protein biosynthesis system including nonnatural amino acids

    Ohtsuki, T, Doi, Y, Manabe, T, Sisido, M

    IEEE International Symposium on Micromechatronics and Human Science (MHS)  2006 

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  • ペプチド核酸(PNA)-膜透過ペプチド(CPP)コンジュゲートを用いたDNAの細胞内導入

    第16回アンチセンスシンポジウム  2006 

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  • Antisense effects of pyrrolidine-based oxy-PNAs conjugated with a membrane-permeable peptide in Escherichia coli

    43JPS-PEM4  2006 

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  • ピロリジン環型オキシPNA(POPNA)の大腸菌内アンチセンス効果

    第16回アンチセンスシンポジウム  2006 

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  • "DNA delivery into CHO cells by the use of complementary peptide nucleic acid conjugated with a cell-penetrating peptide

    NANOBIO-TOKYO2006  2006 

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  • タンパク質への部位特異的なリン酸化セリン導入法の開発

    第21回生体機能関連化学部会・第9回バイオテクノロジー部会・第9回生命化学研究会合同シンポジウム  2006 

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  • 哺乳動物細胞内における蛋白質への位置特異的蛍光基導入

    第21回生体機能関連化学部会・第9回バイオテクノロジー部会・第9回生命化学研究会合同シンポジウム  2006 

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  • 非天然アミノ酸を担持したtRNAに対して結合能の高いEF-Tuの創製

    第21回生体機能関連化学部会・第9回バイオテクノロジー部会・第9回生命化学研究会合同シンポジウム  2006 

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  • RNA isolation using biotinylated PNA.

    20th IUBMB International Congress of Biochemistry and Molecular Biology  2006 

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  • Establishment of purification method of C. elegans mitochondrial ribosome by peptide affinity tags.

    20th IUBMB International Congress of Biochemistry and Molecular Biology  2006 

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  • ISFET型電極を用いるアミノ酸センサーの開発

    日本化学会第86春季年会  2006 

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  • 寄生性線虫を用いた、異常構造をもつミトコンドリアtRNAとEF-Tuの相関関係の解明

    第75回日本寄生虫学会大会  2006 

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  • 非天然アミノ酸含有翻訳システムの拡張

    第8回日本RNA学会年会  2006 

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  • Specific RNA binding of immobilized PNAs

    29th European Peptide Symposium  2006 

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  • Creation of EF-Tu to incorporate large nonnatural amino acids into proteins

    20th IUBMB International Congress of Biochemistry and Molecular Biology  2006 

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  • 細胞膜透過性RNA結合タンパク質によるshRNAの細胞内導入

    第8回日本RNA学会年会  2006 

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  • 固定化PNAによるRNA精製法

    第8回生命化学研究会シンポジウム  2006 

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  • 大腸菌に対する主鎖骨格にピロリジン環を持つオキシペプチド核酸(POPNA)のアンチセンス効果

    日本化学会第86春季年会  2006 

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  • 人工遺伝暗号を用いた非天然アミノ酸導入系の最適化 Invited

    大槻高史

    特定領域「分子計算設計論」公開シンポジウム 企画セミナー「人工遺伝暗号系」  2005.3.11 

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  • RNA isolation using immobilized PNA

    Ohtsuki, T, Kumano, C, Manabe, T, Kitamatsu, M, Sisido, M

    4th International Symposium on Nucleic Acids Chemistry(第32回核酸化学シンポジウム)  2005 

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  • RNA結合蛋白質を介したshRNAの細胞内導入法

    第28回日本分子生物学会年会  2005 

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  • アミノアシルtRNA合成酵素を用いる アミノ酸センシング

    日本化学会第85春季年会  2005 

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  • 縮小化した機能性RNAからなる線形動物ミトコンドリアのタンパク質合成系

    第74回日本寄生虫学会大会  2005 

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  • ミトコンドリアtRNA-m1A9修飾酵素の探索

    第28回日本分子生物学会年会  2005 

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  • C. elegansミトコンドリアリボソームの精製を目指したペプチドタグ付リボソームタンパク質の発現

    第28回日本分子生物学会年会  2005 

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  • 非天然アミノ酸を担持したtRNAに対して結合能の高いEF-Tuの創製

    第28回日本分子生物学会年会  2005 

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  • 異常構造をもつミトコンドリアtRNAとEF-Tuの相関関係の解明

    第28回日本分子生物学会年会  2005 

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  • EF-Tu binding ability of tRNAs carrying various types of nonnatural amino acids

    4th European-Japanese Bioorganic Conference & Chemical Biology COE Program Sponsored by Okayama University  2005 

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  • セリン特異的なEF-Tuの発見とセリン特異性に関与するアミノ酸残基の解析

    第28回日本分子生物学会年会  2005 

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  • 非天然アミノ酸を担持したtRNAとEF-Tuとの結合評価

    第27回日本分子生物学会年会  2004 

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  • 線形動物ミトコンドリアEF-Tu2のtRNA認識機構の解明

    第6回RNAミーティング  2004 

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  • Orthogonal tRNAs for four-base codon translation

    International Symposium on Nucleic Acids, Membranes and Signal Transduction ---Symposium in Honor of Professor H. Gobind Khorana  2004 

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  • 線形動物ミトコンドリアにみられるtRNAとペプチド伸長因子EF-Tuの共進化

    第73回日本寄生虫学会大会  2004 

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  • 無脊椎動物ミトコンドリアのセリルtRNA合成酵素組換えタンパク質の発現

    第27回日本分子生物学会年会  2004 

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  • 4塩基コドンを用いた翻訳のための直交化tRNAの探索

    第27回日本分子生物学会年会  2004 

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  • Elongation factors Tu for truncated tRNAs

    International Symposium on Nucleic Acids, Membranes and Signal Transduction ---Symposium in Honor of Professor H. Gobind Khorana  2004 

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  • Nonnatural amino acid incorporation into protein by mitochondrial tRNAs having non-standard structure

    RNA Structure and Function: a Joint Biochemical Society/Royal Society of Chemistry Focused Meeting  2004 

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  • セリルtRNAのみを認識する伸長因子EF-Tu

    第26回日本分子生物学会年会  2003 

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  • Characteristic features of mammalian mitochondrial translation systems and functional equivalency of mitochondrial tRNA and translation factors to E. coli counterparts

    International symposium commemorating the opening of cell-free science and technology research center  2003 

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  • Decoding property of C5 uridine modification at the wobble position of tRNA anticodon.

    Kurata, S, Ohtsuki, T, Wada, T, Kirino, Y, Takai, K, Saigo, K, Watanabe, K, Suzuki, T

    30th symposium on Nucleic Acids Chemistry  2003 

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  • Unique translation systems of animal mitochondria

    International congress on protein expression & protein function  2003 

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  • Modified nucleotides in nematode mitochondrial tRNAs

    20th International tRNA Workshop  2003 

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  • Unique serine-specificity of mitochondrial EF-Tu

    20th International tRNA Workshop  2003 

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  • Towards artificial repair of UV-damaged DNA: studies on drug binding and alkali hydrolysis.

    Iwai, S, Inase, A, Jafar, S, Higurashi, M, Ohtsuki, T, Xu, Y, Sugiyama, H

    30th symposium on Nucleic Acids Chemistry  2003 

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  • Unusual elongation factors Tu for truncated tRNAs

    20th International tRNA Workshop  2003 

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  • 線形動物ミトコンドリアにみるtRNAとペプチド伸長因子EF-Tuの共進化

    第26回日本分子生物学会年会  2003 

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  • Unique characteristics of animal mitochondrial translation systems

    20th International tRNA Workshop  2003 

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  • 延長部分を持つ伸長因子TuによるTアーム欠失tRNAの新規認識機構

    第26回日本分子生物学会年会  2003 

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  • Two distinct elongation factors Tu specific for respective T arm-lacking and D arm-lacking tRNAs in nematode mitochondria

    19th International tRNA workshop  2002 

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  • Chemical synthesis of novel taurine-containing uridine derivatives.

    Wada, T, Shimazaki, T, Nakagawa, S, Ohtsuki, T, Kurata, S, Suzuki, T, Watanabe, K, Saigo, K

    29th Symposium on Nucleic Acids Chemistry  2002 

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  • Taurine is a constituent of mammalian mitochondrial tRNAs; new insights into the functions of taurine and human mitochondrial diseases.

    19th International tRNA workshop  2002 

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  • Requirement of modified residue m1A9 for EF-Tu binding to nematode mitochondrial tRNA lacking the T arm.

    Sakurai, M, Ohtsuki, T, Watanabe, Y, Watanabe, K

    28th Symposium on Nucleic Acids Chemistry  2001 

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  • 非天然型塩基対を用いた転写・翻訳系の構築 〜6塩基系による非天然型アミノ酸のタンパク質中への取り込み

    第3回 RNA ミーティング  2001 

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  • Structural and functional compensation for deficit in RNA with proteins in animal mitochondrial translation systems

    International conference in honor of Alexander Spirin:PROEIN SYNTHESIS  2001 

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  • Unnatural base pairs between 2-amino-6-(2-thienyl)purine and the complementary bases.

    Hirao, I, Fujiwara, T, Kimoto, M, Mitsui, T, Okuni, T, Ohtsuki, T, Yokoyama, S

    27th Symposium on Nucleic Acids Chemistry  2000 

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  • RNAの残基特異的安定同位体標識法 Invited

    大槻高史

    第9回NMR若手勉強会  1999.6.3 

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  • ホウ素含有化合物およびそれを含む薬剤

    北松瑞生, 副島哲朗, 山形尚紀, 道上宏之, 大槻高史

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    Application no:特願2019- 008817  Date applied:2019.1.22

    Announcement no:特開2020-117479  Date announced:2020.8.6

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  • 修飾RNAを含有する分子集合体及びそれを用いたRNA送達システム

    大槻高史, 松浦栄次, 小渕浩嗣, 赤星彰也, 小関 英一

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    Applicant:株式会社島津製作所・岡山大学

    Application no:特願2016-557827  Date applied:2014.11.8

    Patent/Registration no:特許6586103  Date registered:2019.9.13 

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  • 近赤外光によりRNAを細胞質内に送達するための新規なキャリア分子ならびに方法

    大槻高史, 石躍由佳

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    Application no:特願2011-063953  Date applied:2011.11.24

    Patent/Registration no:特許5900895 

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  • 新規ペプチド複合体、そのハイブリッド複合体およびその用途

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    Application no:特願2011-233812  Date applied:2011.10.25

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  • 乳酸菌により二本鎖RNAを生成するキット及びその利用

    大槻高史, 宍戸昌彦, 公文裕巳, 柏倉祐司, 落合和彦

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    Application no:特願2008-279228  Date applied:2008.10.30

    Patent/Registration no:特許5660537 

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  • 乳酸菌において二本鎖RNAを生成するキット及びその利用

    大槻高史, 宍戸昌彦

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    Application no:特願2008-1174884  Date applied:2008.4.28

    Announcement no:特開2008-301812 

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  • 修飾型PNA/RNA複合体

    北松瑞生, 大槻高史, 宍戸昌彦, 久保貴紀, 大庭英樹

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    Application no:特願2007-37041  Date applied:2007.2.16

    Announcement no:特開2008-301812 

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  • 部位特異的にリン酸化した蛋白質の合成方法、当該方法に用いるホスホセリルtRNA及び当該方法を実施するための試薬キット

    大槻高史, 宍戸昌彦, 横川隆志, 大野敏, 西川一八

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    Application no:特願2006-240828  Date applied:2006.9.5

    Announcement no:特開2008-061538 

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  • 部位特異的アミノ酸導入法のための新規な直交化tRNA

    川井淳, 川上文清, 大槻高史, 宍戸昌彦

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    Application no:特願2005-102868  Date applied:2005.3.31

    Patent/Registration no:特許4406731 

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  • 細胞内へ核酸を導入する為の新規な分子並びに細胞内へ導入する核酸および細胞内へ核酸を導入する為の新規な方法

    川井淳, 川上文清, 大槻高史, 宍戸昌彦

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    Application no:特願2005-103703  Date applied:2005.3.31

    Patent/Registration no:特許4709971 

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  • アミノ酸分析用バイオセンシング装置およびアミノ酸分析用バイオセンサーおよびアミノ酸分析用アミノアシルtRNAシンセターゼ

    釘宮章光, 大槻高史, 菅野憲一

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    Application no:特願2004-359328  Date applied:2004.12.13

    Announcement no:特開2006-166709 

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Awards

  • 研究功績賞

    2017   岡山大学工学部  

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  • 内山勇三科学技術賞

    2016  

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    Country:Japan

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  • The Journal of Biochemistry/OUP Poster Prize

    2009  

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    Country:Japan

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  • 岡山大学若手トップリサーチャー研究奨励賞

    2008  

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    Country:Japan

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  • 生体機能関連化学シンポジウム講演賞

    2007  

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    Country:Japan

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Research Projects

  • ケージド相分離ペプチドからなる相分離液滴の光制御と応用

    Grant number:25K01897  2025.04 - 2028.03

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    大槻 高史

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    Grant amount:\18720000 ( Direct expense: \14400000 、 Indirect expense:\4320000 )

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  • 光制御可能な相分離顆粒デバイスの開発

    Grant number:23H04422  2023.04 - 2025.03

    科研費  学術変革領域研究(A)

    大槻 高史

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    Grant amount:\10400000 ( Direct expense: \8000000 、 Indirect expense:\2400000 )

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  • Photochemical internalization of mRNAs

    Grant number:22K19895  2022.06 - 2024.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Research (Exploratory)

    大槻 高史, 位高 啓史

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    Grant amount:\6500000 ( Direct expense: \5000000 、 Indirect expense:\1500000 )

    mRNAは、遺伝子治療と比べてゲノムへの挿入変異リスクもなく、原理的にどのようなタンパク質でも産生可能であるため、疾患治療に向けて広範な応用の可能性がある。しかしながら、mRNAは生体内での安定性が極めて低く、標的特異的デリバリーが困難である。標的特異的医薬デリバリー戦略の1つに光でターゲティングする方法が挙げられ、その原理としては、薬剤キャリアと光増感剤を用いたphotochemical internalization(PCI)が有力だと考えられる。PCIは、エンドサイトーシスで細胞内に取り込まれてエンドソーム内蓄積する物質を、エンドソーム蓄積性の光増感剤を併用して、光依存的にエンドソームから脱出させ、細胞質内に導入する方法である。本研究では、(1) PCI用の光増感剤と様々な核酸キャリアを組み合わせて、光依存的なmRNAデリバリーを検討した。また、(2) ペプチド基材を用いた光増感剤搭載型の独自キャリアについても試作検討を行い、mRNA運搬が可能なものが見出された。(1)においては興味深いことに、同じ光増感剤を用いても、キャリアによって大きく結果が異なり、(a)光なしでもある程度のmRNA導入が起こるが、光照射時に導入の促進が全く起こらない、(b)光を照射してもしなくてもmRNA導入がほとんど起こらない、(c)光なしではmRNA導入が起こらないが、光照射すると高効率に導入が起こる、などの違いがあった。つまり、(c)により光照射した細胞に特異的な導入に成功した。

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  • mRNAのPhotochemical Internalization

    2022.04 - 2024.03

    科研費 挑戦的研究(萌芽) 

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\5000000

  • 生命機能の光制御のための機能性光増感ペプチドの開発

    2018.04 - 2022.03

    科研費 基盤研究(B) 

    大槻 高史、舟橋 弘晃

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  • Creation of mammalian oocytes with high fertility by spatiotemporal regulation of the function of mitochondria

    Grant number:18H02328  2018.04 - 2022.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    FUNAHASHI Hiroaki

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    Grant amount:\17290000 ( Direct expense: \13300000 、 Indirect expense:\3990000 )

    The aim of this study was to generate oocytes with high developmental competence by regulating the mitochondrial (mt) DNA copy number in the ooplasm. Porcine ooplasmic mtDNA copy number did not change during in vitro maturation (IVM) and was lower in oocytes from small than medium follicles. Drp1 (mt mitogen) was reduced during early development, and its inhibition impaired the mt distribution in oocytes and early developmental competence. Overexpression of PGC1alpha (mt synthesis-related genes) resulted in increased reactive oxygen species and mtDNA copy number reduction in oocytes, and reduced IVM rate and early developmental competence. The autophagic potential of oocytes varied greatly during IVM and early development, and differed significantly by follicle diameter of origin. Attempts to introduce photosensitizers and peptide/protein molecular tools into oocytes have not resulted in significant improvement.

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  • ケージドアミノアシルtRNAによる初期胚における翻訳の時空間的制御

    2017.04 - 2019.03

    科研費 挑戦的研究(萌芽) 

    大槻 高史, 若井 拓哉

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    Authorship:Principal investigator  Grant type:Competitive

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  • Live imaging of microRNA by intracellular-invasion peptide nucleic acid beacon

    Grant number:16KT0195  2016.07 - 2019.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    KITAMATSU Mizuki

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    Grant amount:\4680000 ( Direct expense: \3600000 、 Indirect expense:\1080000 )

    We aim to develop methods to detect microRNAs that are indicators of disease in cells. In particular, dell-penetrating peptide and peptide nucleic acid (PNA) beacon were conjugated for direct detection of microRNA inside cells. We also used the PNA beacon to enable detection of the target microRNA based on the color of the fluorescence emission by modified with two fluorescent groups that generate fluorescence resonance energy transfer at N- and C- terminal on the PNA. We successfully detected the RNA by using the PNA beacon in vitro.

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  • 細胞内におけるRNA分解の観測法の開発とストレス応答研究への応用

    2015.04 - 2017.03

    科研費 挑戦的萌芽研究 

    大槻 高史、渡邉 和則

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    Authorship:Principal investigator  Grant type:Competitive

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  • 光増感剤修飾分子を用いたPCI の分子科学

    2014.04 - 2016.03

    科研費 新学術領域研究(ナノメディシン) 

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

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  • PCDR技術による細胞機能の光制御

    2013.04 - 2016.03

    科研費 基盤研究(B) 

    大槻 高史, 渡邉 和則

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    Authorship:Principal investigator  Grant type:Competitive

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  • Sonochemical internalization法の確立

    2013.04 - 2015.03

    科研費 挑戦的萌芽研究 

    大槻 高史

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  • 光増感によるエンドソーム脱出の分子科学

    2012.04 - 2014.03

    科研費 新学術領域研究(ナノメディシン) 

    大槻 高史

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  • Analyses of cell-cycle dependency of intracellular events induced by RNAs

    Grant number:23651221  2011.04 - 2013.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research

    OHTSUKI Takashi

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    Authorship:Principal investigator  Grant type:Competitive

    Cell cycle dependency of RNAi was analyzed by introducing RNA into cell cycle synchronized cells and by introducing RNA into cells, in which a specific cell cycle was visualized. As a result, cell cycle dependency of RNAi was shown, but reproducibility of the result should be confirmed In addition, to examine cell cycle dependency of cell differentiation induced by RNA, we established a method to promote muscular cell differentiation promotion by introducing the RNA into cells.

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  • 乳酸菌をキャリアとしたRNAiデリバリー

    2010.04 - 2011.03

    JST  研究成果展開事業 A-STEP 探索タイプ 

    大槻 高史

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    乳酸菌を用いてRNAiデリバリーを行う手法の開発に取り組んだ。本手法による癌細胞増殖抑制効果を示すこと、および、腸内デリバリー効果を示すことを目標とした。まず、乳酸菌RNAiデリバリー法により、マウス皮下に植え付けたヒト癌細胞の増殖抑制を試みた。現在、この方法による癌細胞の増殖抑制については実現できていない。また、乳酸菌のマウスへの経口投与による腸内への核酸送達を調べたが、現在のところ明確に送達されたことを示すデータは得られなかった。しかしながら、ルシフェラーゼの発現抑制という形で、乳酸菌RNAiデリバリー法によるRNAi効果は示されたので、今後の研究継続により本技術の実用性が示されることが期待される。

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  • Development of CLIP-RNAi method

    Grant number:21686078  2009.04 - 2012.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (A)

    OHTSUKI Takashi

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    Authorship:Principal investigator  Grant type:Competitive

    RNAi is currently an important experimental tool for basic research into gene function. RNAi also has many potential uses in various biotechnological and therapeutic applications. We recently developed photoinducible RNAi method, termed CLIP-RNAi. In this study, we improved the photoresponsive RNA carrier for CLIP-RNAi. We demonstrate the use of CLIP-RNAi with several fluorescent dyes that can be excited at different wavelengths. We used CLIP-RNAi method in in vitro and in vivo applications.

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  • RNAi創薬を目指したRNAデリバリー技術の開発

    Grant number:20015032  2008.04 - 2010.03

    科研費  特定領域研究(がん研究)

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    最近、研究代表者らは、細胞内侵入性ペプチド(CPP)融合型RNA結合蛋白質(RBP)と、そのRBPの認識RNA配列につながったsiRNAを組み合わせて細胞内に導入する方法を開発した。昨年度からの本助成研究において手法の確立が進んだが、本年度はがん治療への応用を目指して、CPP-RBPによるRNA導入法に関して以下の検討を行った。
    1) CPP-RBPでsiRNAを細胞内導入することにより、ヒトの大腸癌細胞由来株であるSW480株の増殖抑制が可能かどうかを検討した。大腸癌細胞由来株で特異的に発現しているK-Ras変異体遺伝子のRNAi法による発現抑制により、その増殖抑制を試みたが現状では顕著な増殖抑制効果は見られていない。今後のCPP-RBPの改良により明確な効果が現れることを期待している。
    2) 腸内の癌の治療を目指し、乳酸菌にCPP-RBP/shRNAを合成させて腸内送達する方法の開発を目指す。現状ではshRNAのみを産生する乳酸菌株を作製し、マウスにおいてこの乳酸菌を投与することで特異的なRNAi効果が起こることを見いだした。今後CPP-RBPとshRNAの共発現系を内包する乳酸菌を作ることにより、このRNAi効果がより顕著になることを期待している。
    3) 癌細胞で過剰発現がみられる膜蛋白質を標的とした抗体あるいはペプチドを、CPP-RBPに融合させることにより、CPP-RBP/shRNA複合体が癌細胞にのみ結合しやすくなり、それに伴いshRNAの細胞内導入効率RNAi効果が高まらないかと考えた。本年度はその例として卵巣癌や乳癌などで過剰発現がみられるErbB2に結合する蛋白質をCPP-RBPに融合させたものを作製した。現在、これにより通常のCPP-RBPと比べて癌細胞の増殖抑制効果が高まるかどうかを調べている。

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  • 拡張型蛋白質合成系を含む細胞システムの構築

    Grant number:20034036  2008.04 - 2010.03

    科研費  特定領域研究(バイオ操作)

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    本課題では、拡張型蛋白質合成系(蛋白質へ非天然アミノ酸を導入する系)を含む細胞システムを構築すること、および、拡張型蛋白質合成系を含む人工細胞システムを構築することを目的とする。本年度は、以下について研究を行った。
    1) 細胞内で非天然アミノ酸を含む蛋白質を合成するシステムの開発:細胞内で非天然アミノ酸を含む蛋白質の合成を行うため、アミノアシルtRNA(ここでは、非天然アミノ酸を担持したtRNA)の細胞内導入法を検討した。リポフェクション、マイクロインジェクション、およびエレクトロポレーション法でアミノアシルtRNAは細胞内に導入できるものの、細胞内での蛋白質合成には極めて低い効率でしか使われないことが分かってきた。この理由は、(1)アミノアシルtRNAの分解が速いため(導入操作中に、または、導入操作でダメージを受けた細胞が活発な蛋白質合成を再開するまでに、分解してしまうため)か、(2)細胞内の翻訳システムにフリーのアミノアシルtRNAが入り込むのが難しいためだと考えている。現在、(1)に対しては、アミノアシルtRNAの化学的保護および蛋白質による保護、(2)に対しては、アミノアシルtRNAと翻訳因子eEF1Aの複合体を細胞内導入すること、などの対策を行っているところである。
    2) 人工細胞内で非天然アミノ酸を含む蛋白質を合成するシステムの開発:人工モデル細胞(リポソーム)中に無細胞蛋白質合成系を内包して、人工細胞内で蛋白質合成を行うようなシステムが報告されている。このシステムに、4塩基コドンを持つmRNAと非天然アミノ酸を担持したアミノアシルtRNAを加え、4塩基コドンで指定した部位に非天然アミノ酸を含む蛋白質の合成が可能であることを示した。このシステムで膜蛋白質ロドプシンに蛍光性非天然アミノ酸を導入することに成功し、これが膜に局在することも分かってきた。

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  • 人工核酸によるRNAの検出と精製

    Grant number:19021034  2007.04 - 2009.03

    科研費  特定領域研究(ライフサーベイヤ)

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    人工核酸の一種であるペプチド核酸(PNA)は、ペプチド骨格に核酸塩基が付いた構造をもつ。PNAの特徴の一つは、相補的なRNAと非常に強く結合することであり、同じ配列のDNAやRNAと比べても格段にRNA結合能が高く、また配列特異性も優れている。本研究では、この特性を生かして、以下の2つの研究を行った。
    1) 近年miRNAと呼ばれる小さなRNAが多数存在し、広範な生命機能を制御していることが明らかになり、様々な状態における生体内のmiRNA発現量を調べることが生命科学研究における重要課題となってきた。本研究課題では、PNAがRNAに特異的に強く結合する特性を生かし、miRNAを高感度に検出するためのPNAアレイの開発を行った。様々な状態の細胞、特に様々な疾患におけるmiRNAの発現量を網羅的に調べることは、現在急務になっている。そのため疾患に関係するmiRNAなどを題材としてPNAアレイの能力を検証した。10merのPNAプローブを用いたところ, miRNAの検出によく用いられるDNAプローブ(10merおよび20mer)と比べ, 非常に感度の良い検出が可能になった。
    2) RNA混合物の中から単一のRNA種を精製する方法として、PNAを固相担体とする簡便なRNA精製法の開発に取り組んだ。固定化PNA法が、低温で高効率にRNAを単離でき、多くのRNAに拡張できる方法であることを示した。

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  • New development in study of mitochondrial translation system of theparasitic nematode

    Grant number:19590425  2007.04 - 2009.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    YOSHINARI Shigeo, WATANABE Yoh-ichi, OHTSUKI Takashi, YOKOBORI Shin-ichi

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    Authorship:Coinvestigator(s) 

    Grant amount:\4550000 ( Direct expense: \3500000 、 Indirect expense:\1050000 )

    動物ミトコンドリア(mt)のタンパク質合成系は、より短い機能性RNAを用いる。我々は、短縮化したmt tRNA を持つ種と持たない種が混在する節足動物で、短縮化したtRNAを持たない昆虫の2つのEF-Tu のうちの1つが、短縮化したtRNA と結合できることを示した。また、植物寄生性線虫ミトコンドリアで遺伝子が未同定だったtRNA のうち1種の発現を同定した。このtRNA は発現が確認された中で最も短い。

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  • 多様な非天然アミノ酸が導入可能なタンパク質合成系の創製

    2007.04 - 2008.03

    JST  研究成果展開事業 シーズ発掘試験 

    大槻 高史

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    Authorship:Principal investigator 

    蛋白質の特定部位に非天然のアミノ酸を導入することができれば、創薬および蛋白質研究に対して多大な貢献が見込まれる。既存の非天然アミノ酸導入システムでは、蛋白質への導入が不可能な非天然アミノ酸が多数存在する。その原因の一つは非天然アミノ酸がEF-Tuという因子と適合しないことであった。本研究では、多様な非天然アミノ酸の蛋白質への導入を可能にすることを目的とし、EF-Tuを改変して非天然アミノ酸の導入を検討する。

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  • 非天然アミノ酸含有タンパク質の無細胞及び細胞モデル系での発現

    Grant number:18048031  2006.04 - 2008.03

    科研費  特定領域研究(バイオ操作)

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    生命機能の再構成という目において、蛋白質生合成系の役割は大きい。本研究では、蛋白質へ非天然アミノ酸を導入する技術の開発を通じて、蛋白質合成の分子基盤の理解と、人工的な細胞機能の再構築を目指す。具体的には、非天然アミノ酸含有蛋白質合成系をtRNAや翻訳因子の改良により更に効率化し、無細胞系及びモデル細胞系で利用することを目的とする。本年度は、以下の項目に取り組んだ。
    (1)改良した合成システムによる新規な人工蛋白質の創製
    前年度に改変を行ったEF-Tu、及び、開発した4塩基コドン用tRNAにより、以下の(i).(ii)を試みた。
    (i)従来取り込み効率の低かった非天然アミノ酸の蛋白質への導入:改変EF-Tuを用いることにより これまでほとんど蛋白質へ導入できなかった1-pyrenylalanineの導入が可能になったという結果を前年度に得ている。このような例を更に数例調べ、導入効率の極めて低かった2-anthraquinonylalanineおよび1-pyrenylalanineの効率的な導入が可能になったことを明らかにした。これらの結果および前年度の結果は本年度11月にJ.Am. Chem.Soc誌に掲載された。
    (ii)蛋白質への複数種の非天然アミノ酸の同時導入:4塩基コドン用に開発した高性能なtRNAを用いて1つの蛋白質に複数種の非天然アミノ酸の同時導入を試みた。その結果、3種類の非天然アミノ酸の同時導入が可能になった。
    (2)細胞中での非天然アミノ酸含有蛋白質発現
    上の高効率非天然蛋白質発現系を細胞内に導入し、その中で蛍光ラベル蛋白質を合成させることを目指した。細胞としては哺乳動物細胞を用いた。現在のところ、細胞内への蛍光アミノアシルtRNAの導入法の開発途上であり、わずかながら導入できることが認められた。今後さらに導入法を最適化し、蛍光アミノアシル化tRNAが細胞内翻訳系で用いられて蛍光ラベル蛋白質の産生に結びつくことを確認したい。

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  • miRNA発現量解析のためのペプチド核酸アレイの開発

    2006.04 - 2007.03

    JST  研究成果展開事業 シーズ発掘試験 

    大槻 高史

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    近年miRNA と呼ばれる小さなRNA が広範な生命機能を制御していることが明らかになり、様々な状態における生体内のmiRNA 発現量を調べることがライフサイエンス研究における重要課題となってきた。本課題では、ペプチド核酸(PNA)の RNA に特異的に強く結合する特性を生かし、miRNA を高感度に検出するためのPNA アレイの開発を行う。先行技術であるDNA アレイと比較して、PNA アレイがmiRNA の検出感度や特異性の上で優れていることが証明できればmiRNA 解析チップとして実用化が可能になる。

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  • RNAの新規細胞内導入法の開発と応用

    2006.01 - 2008.12

    NEDO  産業技術研究助成事業 

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\48100000

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  • 生細胞中での蛋白質の部位特異的蛍光標識法の開発

    Grant number:17750162  2005.04 - 2007.03

    科研費  若手研究(B)

    大槻 高史

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    細胞内の生体分子の動態や局在などのネットワークを調べることはポストゲノム時代の最重要課題の1つである。この課題を網羅的・効率的に進めるためには、生体分子を可視化させる蛍光標識技術の開発が必要である。蛋白質分子の蛍光標識法としては、目的蛋白質の末端にGFPなど蛍光蛋白質を結合させる方法が現在一般的であるが、大きな蛍光蛋白質を融合することにより目的蛋白質の本来の性質が損なわれる可能性も高い。そこで、蛋白質の機能に極力影響を与えない小さな蛍光基の導入が望まれる。本課題では、細胞内で蛋白質中の望んだ部位に蛍光基を導入する方法を開発することを試みた。本方法は細胞内の蛋白質合成装置を利用し、蛍光基をもつアミノ酸を直交化tRNAに結合させ、mRNA上のコドン特異的に蛋白質に導入するというものである。この方法では、蛍光部位は小さな蛍光基としてアミノ酸一残基分のスペースに導入するため、蛋白質の機能を損なう可能性は少ない。
    本年度は、細胞内翻訳装置により蛍光アミノ酸を蛋白質に組み込む方法を検討した。蛍光アミノ酸に対応するコドンとしてアンバーコドンを用い、アンバーコドンを含む蛋白質遺伝子(ここではDsRed遺伝子)を発現ベクターに組み込んで、哺乳動物細胞(ここではCHO細胞)に導入した。その細胞に細胞内のARSによりアミノアシル化されるようなサプレッサーtRNAをリポフェクション法により導入したところ、DsRedの赤色蛍光によりアンバーコドンのサプレッションが観測された。細胞外で蛍光アミノ酸(Bodipy-FL-AF)を結合させたサプレッサーtRNAを用いた場合も、アミノアシルtRNAのエンドソーム経由での細胞内導入が確認された。主にエンドソーム内にアミノアシルtRNAが固まって強い蛍光を放っていたため、これを改善するような導入法を今後さらに検討する。

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  • Study of parasite mitochondrial translation system which have truncated functional RNAs

    Grant number:17590368  2005.04 - 2007.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    WATANABE Yoh-ichi, OHTSUKI Takashi

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    Grant amount:\3600000 ( Direct expense: \3600000 )

    Translation system in metazoan mitochondria is quite different from those in eukaryotic cytoplasm and prokaryote in many points. Among those, based on mitochondrial DNA sequences, translation system in nematode mitochondria has been predicted to have shortest tRNAs and shortest rRNA. From the results of analysis of mitochondrial translation system of parasitic and free-living nematodes, we are hoping to reveal that what will happen when functional RNAs in translation system is truncated. In this study, we analyzed these issues and pursued to reveal why truncation of functional RNAs in translation system had happened. We obtained the following results during the last two years:
    1.To purify a larger amount of mitochondrial ribosome more efficiently, we prepared recombinant nematode expressing a mitochondrial ribosomal protein with affinity tags. However, we could not obtained mitochondrial ribosome from the transformant, suggesting that optimization of combination of ribosomal protein and tag sequence will be necessary.
    2.Using mutant thermophilic bacterial EF-Tu proteins possessing residues found in Caenorhabditis elegans mitochondrial EF-Tu2, we revealed which residues in EF-Tu2 contribute the specificity against seryl-moiety in seryl-tRNA binding.
    3.We revealed specificities of aminoacyl-tRNA binding of mitochondrial EF-Tu proteins in parasitic nematode Trichinella spp.
    4.We revealed specificities of aminoacyl-tRNA binding of mitochondrial EF-Tu proteins in the arthropod.
    5.We found a candidate tRNA methyltransferase.

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  • リン酸化セリンを含む蛋白質作製法の開発

    2005.04 - 2006.03

    JST  研究成果展開事業 シーズ発掘試験 

    大槻 高史

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    蛋白質の働きは、種々の翻訳後修飾により制御されており、その中でもリン酸化修飾はシグナル伝達や転写制御において非常に重要な役割をしている。様々なリン酸化蛋白質は、それぞれについて異なるリン酸化酵素により翻訳後修飾されることが知られている。しかしながら、様々なリン酸化蛋白質の生化学的解析やそれらを用いた臨床研究のためには、個別なリン酸化方法をとるのは煩わしく、一般的な作製方法の開発が望まれる。2005年、古細菌においてリン酸化セリンをtRNAに結合させる酵素(SepRS)が発見されたことに基づき、本研究では、翻訳後修飾ではなく、翻訳中にリン酸化セリンを直接蛋白質に導入する方法(左図参照)を開発する。現在、平成17~18年度の科研費若手Bの助成を受けているが、本課題とは異なる内容である。

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  • Chemical Expansion of the Central Dogma towards Synthetic Microorganisms

    Grant number:15101008  2003.04 - 2008.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (S)

    SISIDO Masahiko, OHTSUKI Takashi, TAKI Masumi

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    Grant amount:\103610000 ( Direct expense: \79700000 、 Indirect expense:\23910000 )

    This research project aims to expand molecules like amino acids, nucleic acids and proteins constitute the protein biosynthesizing system to synthesize novel type of proteins with artificial functions. The final goal is to create synthetic microorganisms that live with 21 type of amino acids. During the 5 year we obtained the following results. (1) Some nonnatural amino acids were successfully charged onto a specific tRNA by the use of a peptide nucleic acid (PNA) that is complementary to the tRNA. The PNA-assisted aminoacylation mold be carried out in an E.coli in vitro protein biosynthesizing system to create a protein incorporated with the nonnatural amino add. (2) tRNAs and an elongation faint Tu. (EF-Tu) were expanded to bring large-sized nonnatural amino adds into E.col ribosome and subsequently to a protein. (3) An orthogonal set of aminoacyl tRNAsynthetase (ARS) and tRNA was screened to bring some nonnatural amino acids into proteins in living mammalian cells. (4) Finally these expanded molecular systems were introduced into mammalian cells and the synthesis of proteins containing nonnatural amino acids was detected, although its quantity was very small.
    In the last year, an attempt was made to apply these techniques to attach fluorescent amino acids to peptides and proteins. The fluorescently labeled peptides were screened to dimmer those that bind to cancer cells or to cancer-cell specific proteins.

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  • Study of nematode mitochondrial translation system using parasitic nematodes

    Grant number:15590365  2003.04 - 2005.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    WATANABE Yoh-ichi, OHTSUKI Takashi, KITA Kiyoshi, KAMIYA Haruo

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    Grant amount:\3600000 ( Direct expense: \3600000 )

    Most nematode mitochondria have D-armless tRNAs, which also exist in most metazoan mitochondria, and T-armless tRNAs. On the other hand, bacterial translation elongation factor Tu (EF-Tu) needs T-arm of aminoacyl-tRNA for the complex formation. In order to investigate how nematode mitochondrial EF-Tu recognizes the T-armless tRNA, we cloned EF-Tus from various nemaotdes. Nematode have two distinct EF-Tus. Two EF-Tus, namely EF-Tu1 and EF-Tu2, have novel C-terminal extensions (about 40-60 residues and about 15 residues, respectively) when compared with bacterial EF-Tu. Caernarhabditis elegans and Trichinella britovi EF-Tu1 bound to the T-armless tRNAs, while EF-Tu2 bound to D-armless tRNAs. C.elegans EF-Tus and T.britovi EF-Tu2 could not bind to conventional tRNAs, while T.britovi EF-Tu1 could bind to conventional tRNAs. These results consist well with the fact that C.elegans mitochondria do not have any conventional tRNA but Trichinella mitochondria have. Deletion of 10 residues of C-terminus of both of C.elegans EF-Tus lead to loss of binding to the D-armless or the T-armless tRNAs, suggesting the novel C-Terminal extensions may involve in interaction between EF-Tus and the truncated nematode tRNAs. In summary, the nematode mitochondrial EF-Tu adapted to the deletion of D-arm or T-arm of tRNAs by the gene duplication and aquirment of novel C-terminal extension to make a new interaction between the truncated tRNA and EF-Tu.

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  • Elucidation of basic principle in genetic information translation system by using the specialty of animal mitochondria

    Grant number:14208077  2002.04 - 2004.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (A)

    WATANABE Kimitsuna, SUZUKI Tsutomu, OHTSUKI Takashi

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    Authorship:Coinvestigator(s) 

    Grant amount:\48880000 ( Direct expense: \37600000 、 Indirect expense:\11280000 )

    The aim of this project is to elucidate the basic principle in genetic information translation system by using the specialty of animal mitochondria. This can be done by combining the well-studied E.coli translation system with the special translation system of animal mitochondria. (1)The minimal prerequisite structure of tRNA : The higher-order structure of nematode mitochondrial (mt) tRNA^<Ser>_<UCU>, which is the smallest tRNA in the extant organisms (54 nucleotides, having no D arm and small T arm consisting of 3 base pairs and a loop with 4 bases), was examined by using NMR. It turned out that almost all the base-pairs predicted from its clover-leaf structure existed in solution and the connector region consisting of the truncated D loop and extra loop had a rather flexible structure. This mobility is indispensable for the tRNA^<Ser>_<UCU> to maintain the relative arrangement of the 3'-terminus and the anticodon equal to that of the usual tRNAs. (2)The minimal prerequisite structure of ribosome : In the animal mt ribosome, the content of rRNA is about a half, but that of proteins is doubled, as compared with the counterparts of E.coli ribosome. The functional regions of mt rRNA are strictly conserved in the central parts of two ribosomal subunits and the peripheral regions of rRNA lacking in the mt ribosome are embedded by proteins so as to maintain the whole shapes of ribosomes almost equal between mt and E.coli. It turned out that some of the stem/loop structures in the peripheral regions of mt ribosome are dispensable. (3)What is the trigger in the hydrolysis reaction of GTP which is an energy source in the translation system : It was suggested that the trigger in the hydrolysis reaction of GTP is L7/L12 protein in 50S ribosomal subunit, on the basis of the fact that GTPase activity of EF-G is activated by L7/L12. We have confirmed this proposal by elucidating that E.coli EF-G can function on the mt ribosome in which only the L7/L12 protein is replaced by the counterpart of E.coli ribosome, although E.coli EF-G cannot work on the native mt ribosome.

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  • 異常tRNAを用いた翻訳系

    2002.04 - 2003.10

    科学技術振興調整費 

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\34000000 ( Direct expense: \29130000 )

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  • 伸長因子EF-TuによるTアームを欠くtRNAの認識機構

    Grant number:13780481  2001.04 - 2002.03

    科研費  若手研究(B)

    大槻 高史

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    Authorship:Principal investigator  Grant type:Competitive

    細菌のEF-Tuの研究から、EF-TuはアミノアシルtRNAのアクセプターアームとTアームを認識していることが知られている。線形動物ミトコンドリアにおいてはEF-Tuとの相互作用に必要と思われるTアームを欠失したtRNAが存在している。線形動物ミトコンドリアでは22種類存在するtRNAのうち、20種類がTアームの欠けたtRNAで、残る2種類はDアームを欠失したtRNAであり、それぞれEF-Tu1とEF-Tu2という2種類のEF-Tuにより認識されることが分かっている。本研究では、これらのtRNAのEF-Tuによる認識機構を調べることにより、RNA構造の短縮化が蛋白質によってどのように補われているかを考察することを目的としている。前年度は、EF-Tu1によるTアームを欠いたtRNAの認識機構の解明に取り組み、EF-Tu1は通常と異なるC末端延長部分により、標準的なEF-Tuが結合する部位以外にtRNAL字構造の内側部分を認識していることを明らかにした。本年度は、以上をふまえて、更に研究を進めた。
    1)EF-Tu1とtRNAの相互作用を分子レベルで明らかにするため、EF-Tu1のC-末端部分(約150残基)のNMRによる立体構造解析に取り組み、解析に必要なスペクトルを揃えるところまで到達した。
    2)EF-Tu2によるDアームの欠けたtRNAの認識について調べた。全く同じtRNA部分を持つセリルtRNA、アラニルtRNA、バリルtRNAを用意し、これらに対する結合を調べたところ、EF-Tu2はセリルtRNAのみを認識することがわかった。通常のEF-Tuは全てのアミノアシルtRNAと結合するため、セリン特異的な認識は極めて珍しい例である。線形動物ミトコンドリアではDアームの欠けたtRNAはセリンtRNAのみであるため、EF-Tu2はこれに対応したものと思われる。
    以上のように、Tアームの欠けたtRNAには、新しいtRNA認識部位をもつEF-Tuが対応し、Dアームの欠けたtRNAには、アミノ酸特異的認識を行うEF-Tuが対応していることが明らかになった。動物ミトコンドリアには通常のtRNAと異なる多様なtRNAが存在し、それらがどのように機能しているかは謎であったが、本研究で明らかにしたようにEF-Tuが共進化してtRNA構造の欠損を補っていることが、一つの答えだと考えられる。

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  • Elucidation of the construction principle of the mitochondria translation system by means of structural biology

    Grant number:11308024  1999.04 - 2002.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (A)

    WATANABE Kimitsuna, OHTSUKI Takashi, SUZUKI Tsutomu

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    Grant amount:\34720000 ( Direct expense: \32800000 、 Indirect expense:\1920000 )

    The aim of this research is to elucidate the construction principle of the mitochondria! translation system, by analyzing the functional structures of each component of the animal mitochondrial translation system using X-ray analysis and NMR spectroscopy, focusing on the functional characteristics and structural abnormalities of the components. The following results have been obtained for these three years.
    (1) Bovine mitochondrial in vitro translation system has been established by optimizing concentration of each component and reaction conditions, which has similar efficiency to that of 」. co// system in terms of poly(U)-dependent poly(Phe) synthesis.
    (2) Ribosomes are exchangeable between E. coli and mitochondrial systems, as far as tRNA and translation factors of the same origins are used, and EF-Tu of E. coli origin is used. Thus, the ribosomes are functionally equivalent between E. coli and mitochondrial systems.
    (3) It was verified that 2 species of serine tRNAs with unusual secondary structures can be functional. The low efficiency of the D arm-lacking serine tRNA is derived from a defect in its binding to the ribosomal A site.
    (4) These two serine tRNAs were recognized by a single seryl-tRNA synthetase (serRS) and its recognition sites on the two tRNAs were identified.
    (5) A unique EF-Tu was found which recognized almost all the T arm-lacking tRNAs in nematode mitochondria. It had 57 amino acid extension at the C terminus which was considered to complement the T arm-lacking part of tRNAs. Another EF-Tu recognizing two D arm-lacking serine tRNAs was specific for serine.
    (6) Crystallization of these translation factors are being under way. We have obtained a needle-like srystal for SerRS. The X-ray analysis will be made in future.

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  • NMRによるミトコンドリアtRNAの立体構造解析

    Grant number:97J07014  1998.04 - 1998.12

    日本学術振興会  科研費  特別研究員奨励費

    大槻 高史

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    Grant amount:\900000 ( Direct expense: \900000 )

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  • Research Works for Interdisciplinary Medical Sciences and Engineering (2021academic year) Year-round  - その他

  • Basic of Applied Chemistry and Biotechnology (2021academic year) 1st semester  - 金1~2

  • Chemical biology (2021academic year) Late  - その他

  • Technical Writing and Presentation (2021academic year) Late  - その他

  • Technical Writing and Presentation (2021academic year) Late  - その他

  • Physical Chemistry 3 (2021academic year) 1st semester  - 火5,火6,金1,金2

  • Physical Chemistry 4 (2021academic year) 1st semester  - 火5,火6,金1,金2

  • Biochemistry 3 (2021academic year) Third semester  - 月3,月4,水1,水2

  • Biochemistry 4 (2021academic year) Third semester  - 月3,月4,水1,水2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2021academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology 1 (2021academic year) Third semester  - 金1,金2

  • Basic Biology 1 (2021academic year) Third semester  - 金1,金2

  • Basic Biology 1 (2021academic year) Third semester  - 金1,金2

  • Basic Biology 2 (2021academic year) Fourth semester  - 金1,金2

  • Basic Biology 2 (2021academic year) Fourth semester  - 金1,金2

  • Basic Biology 2 (2021academic year) Fourth semester  - 金1,金2

  • Gene Engineering (2021academic year) Fourth semester  - 火5,火6

  • Gene Engineering (2021academic year) Fourth semester  - 火5,火6

  • RNA technology (2021academic year) Prophase  - 火5~6

  • General Biotechnology and Drug Discovery Technology (2020academic year) Prophase  - 木1,木2

  • Internship for Interdisciplinary Medical Sciences and Engineering (2020academic year) Year-round  - その他

  • Technical English for Interdisciplinary Medical Sciences and Engineering (2020academic year) Late  - その他

  • Research Works for Interdisciplinary Medical Sciences and Engineering (2020academic year) Year-round  - その他

  • Chemical biology (2020academic year) Late  - その他

  • Technical Writing and Presentation (2020academic year) Late  - その他

  • Radiation Safety Usage Engineering 1 (2020academic year) 1st semester  - 金5,金6

  • Radiation Safety Usage Engineering 2 (2020academic year) Second semester  - 金5,金6

  • Radiation Safety Engineering and Experiments (2020academic year) 1st and 2nd semester  - 金5,金6

  • Physical Chemistry 3 (2020academic year) 1st semester  - 火5,火6,金1,金2

  • Physical Chemistry 4 (2020academic year) 1st semester  - 火5,火6,金1,金2

  • Biochemistry 3 (2020academic year) Third semester  - 月3,月4,水1,水2

  • Biochemistry 4 (2020academic year) Third semester  - 月3,月4,水1,水2

  • Basic Biology (2020academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2020academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology (2020academic year) 3rd and 4th semester  - 金1,金2

  • Basic Biology 1 (2020academic year) Third semester  - 金1,金2

  • Basic Biology 1 (2020academic year) Third semester  - 金1,金2

  • Basic Biology 1 (2020academic year) Third semester  - 金1,金2

  • Basic Biology 2 (2020academic year) Fourth semester  - 金1,金2

  • Basic Biology 2 (2020academic year) Fourth semester  - 金1,金2

  • Basic Biology 2 (2020academic year) Fourth semester  - 金1,金2

  • Gene Engineering (2020academic year) Fourth semester  - 火5,火6

  • Gene Engineering (2020academic year) Fourth semester  - 火5,火6

▼display all

 

Social Activities

  • 高等学校理科研修講座(生物)

    Role(s):Lecturer

    岡山県総合教育センター  2019.8.23

  • 専門日本語教育@中国 東北師範大学

    Role(s):Lecturer, Organizing member

    文部科学省  文部科学省中国赴日本国留学生(進学博士)予備教育  2019.7.25 - 2019.9.1

  • 専門日本語教育@中国 東北師範大学

    Role(s):Lecturer, Organizing member

    文部科学省  文部科学省中国赴日本国留学生(進学博士)予備教育  2018.7.25 - 2018.9.3

Media Coverage

  • 遺伝情報の伝達担うRNA、光当てて細胞内に導入 Newspaper, magazine

    山陽新聞  2019.1.28

     More details

    Author:Other 

  • 「RNA干渉」細胞傷めず Newspaper, magazine

    日経産業新聞  2010.1.15

     More details

    Author:Other