2021/10/21 更新

写真a

オオツキ タカシ
大槻 高史
OHTSUKI Takashi
所属
ヘルスシステム統合科学学域 教授
職名
教授
外部リンク

学位

  • 学士 ( 東京工業大学 )

  • 修士 ( 東京大学 )

  • 博士(工学) ( 東京大学 )

研究キーワード

  • 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

  • nonnatural amino acid

  • tRNA

  • translation

  • EF-Tu

  • RNA delivery

  • siRNA

  • Biochemistry

  • Molecular Biology

  • siRNA

  • 光増感剤

  • RNA工学

  • RNAi

  • ケージド化合物

  • RNAi

  • EF-Tu

  • tRNA

  • タンパク質生合成系

  • 非天然アミノ酸

研究分野

  • ナノテク・材料 / 生体化学

  • ライフサイエンス / 構造生物化学

  • ものづくり技術(機械・電気電子・化学工学) / バイオ機能応用、バイオプロセス工学

  • ナノテク・材料 / 生物分子化学

  • ライフサイエンス / 分子生物学

  • ライフサイエンス / 生体医工学

  • ライフサイエンス / 生体材料学

▼全件表示

学歴

  • 東京大学    

    1995年 - 1998年

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  • 東京大学    

    1993年 - 1995年

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    国名: 日本国

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  • 東京工業大学   工学部   生体分子工学科

    1989年 - 1993年

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    国名: 日本国

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経歴

  • 岡山大学   ヘルスシステム統合科学研究科   教授

    2018年4月 - 現在

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  • 岡山大学   自然科学研究科   教授

    2010年6月 - 2018年3月

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  • 岡山大学   自然科学研究科   准教授

    2007年10月 - 2010年5月

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  • 岡山大学   工学部生物機能工学科   講師

    2003年11月 - 2007年9月

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  • 東京大学   大学院新領域創成科学研究科   助手

    2002年4月 - 2003年10月

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  • 東京大学   大学院工学系研究科   助手

    2000年3月 - 2002年3月

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  • 理化学研究所   ゲノム化学総合研究センター   研究員

    1999年1月 - 2000年3月

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  • 日本学術振興会   日本学術振興会特別研究員

    1997年4月 - 1998年12月

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▼全件表示

所属学協会

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委員歴

  • 日本化学会中国四国支部   役員  

    2021年4月 - 現在   

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  • 日本核酸化学会   評議員  

    2016年 - 現在   

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    団体区分:学協会

    日本核酸化学会

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  • 日本生化学会   評議員  

    2011年 - 現在   

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    団体区分:学協会

    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年 - 現在   

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    団体区分:学協会

    日本化学会 生体機能関連化学部会

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論文

  • Fluorophore–PNA–Quencher/Quencher–DNA Probe for miRNA Detection 査読

    Tabara, K., Watanabe, K., Shigeto, H., Yamamura, S., Kishi, T., Kitamatsu, M., Ohtsuki, T.

    Bioorganic & Medicinal Chemistry Letters   in press   2021年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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 査読

    Watanabe K, Nawachi T, Okutani R, Ohtsuki T.

    Sci Rep.   11 ( 1 )   14936 - 14936   2021年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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

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  • Lactosome-Conjugated siRNA Nanoparticles for Photo-Enhanced Gene Silencing in Cancer Cells 査読

    Lim MSH, Nishiyama Y, Ohtsuki T, Watanabe K, Kobuchi H, Kobayashi K, Matsuura E.

    J Pharm Sci.   110 ( 4 )   1788 - 1798   2021年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.xphs.2021.01.026

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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年4月

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    記述言語:英語   出版者・発行元:MDPI  

    DOI: 10.3390/app11083594

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  • Minimization of apoptosis-inducing CPP-Bim peptide. 査読

    Zhou S, Watanabe K, Koide S, Kitamatsu M, Ohtsuki T.

    Bioorg Med Chem Lett   36   127811 - 127811   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.bmcl.2021.127811

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

    Watanabe K, Ohtsuki T.

    Int J Mol Sci.   22 ( 9 )   4982 - 4982   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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. 国際誌

    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年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    "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. 査読

    Kitamatsu, M. Yuasa, H., Ohtsuki, T., Michiue, H

    Bioorganic & Medicinal Chemistry   33   116036 - 116036   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.bmc.2021.116036

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  • Fluorescence lifetime probes for detection of RNA degradation. 査読

    Hirata, R., Hirakawa, K., Shimada, N., Watanabe, K., and Ohtsuki, T.

    Analyst   146   277 - 282   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/d0an01230k

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  • Photoinduced Endosomal Escape Mechanism: A View from Photochemical Internalization Mediated by CPP-Photosensitizer Conjugates. 国際誌

    Tet Htut Soe, Kazunori Watanabe, Takashi Ohtsuki

    Molecules   26 ( 1 )   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Endosomal escape in cell-penetrating peptide (CPP)-based drug/macromolecule delivery systems is frequently insufficient. The CPP-fused molecules tend to remain trapped inside endosomes and end up being degraded rather than delivered into the cytosol. One of the methods for endosomal escape of CPP-fused molecules is photochemical internalization (PCI), which is based on the use of light and a photosensitizer and relies on photoinduced endosomal membrane destabilization to release the cargo molecule. Currently, it remains unclear how this delivery strategy behaves after photostimulation. Recent findings, including our studies using CPP-cargo-photosensitizer conjugates, have shed light on the photoinduced endosomal escape mechanism. In this review, we discuss the structural design of CPP-photosensitizer and CPP-cargo-photosensitizer conjugates, and the PCI mechanism underlying their application.

    DOI: 10.3390/molecules26010036

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  • Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate. 査読

    Kim, H., Watanabe, S., Kitamatsu, M., Watanabe, K., and Ohtsuki, T.

    Sci. Rep.   10   19087 - 19087   2020年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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. 査読 国際誌

    Shigeto H, Yamada E, Kitamatsu M, Ohtsuki T, Iizuka A, Akiyama Y, Yamamura, S.

    Micromachines   11 ( 7 )   E628 - E628   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.3390/mi11070628

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  • Endosomal Escape of Peptide-Photosensitizer Conjugates Is Affected by Amino Acid Sequences near the Photosensitizer 査読 国際誌

    Miyoshi Y, Kadono M, Okazaki S, Nishimura A, Kitamatsu M, Watanabe K, Ohtsuki T.

    Bioconjug Chem.   31 ( 3 )   916 - 922   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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 査読

    Hakata Y, Ishikawa S, Ohtsuki T, Miyazawa M, Kitamatsu M.

    Org Biomol Chem.   18 ( 10 )   1978 - 1986   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/c9ob02559f

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

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

    Analyst   144 ( 15 )   4613 - 4621   2019年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/c9an00725c

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  • A leucine zipper-based peptide hybrid delivers functional Nanog protein inside the cell nucleus. 査読 国際誌

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

    Bioorganic & medicinal chemistry letters   29 ( 7 )   878 - 881   2019年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We synthesized a pair of compounds containing leucine zipper peptides to deliver protein cargo into cells. One is a cell-penetrating peptide (CPP) with Lz(E), a leucine zipper peptide containing negatively charged amino acids, and the other is a Nanog protein with Lz(K), a leucine zipper peptide containing positively charged amino acids. When cells were treated with these equimolar mixtures, Nanog-Lz(K) hybridized with Lz(E)-CPP was successfully delivered into the cells. Furthermore, Nanog-Lz(K) exerted its proper function after nuclear transport.

    DOI: 10.1016/j.bmcl.2019.02.004

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

    Hyungjin Kim, Mizuki Kitamatsu, Takashi Ohtsuki

    J. Biosci. Bioeng.   128 ( 1 )   110 - 116   2019年1月

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    掲載種別:研究論文(学術雑誌)  

    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. 査読

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

    PLOS ONE   14 ( 1 )   e0211505   2019年

  • Red and near-infrared light-directed cytosolic delivery of two different RNAs using photosensitive RNA carriers. 査読

    Shiraga, K, Soe, T. H, Matsumoto, S, Watanabe, K, Ohtsuki, T

    Bioconjugate Chemistry   29 ( 9 )   3174 - 3179   2018年8月

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    記述言語:英語  

    DOI: 10.1021/acs.bioconjchem.8b00487

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

    Kazunori Watanabe, Ryuhei Yamaji, Takashi Ohtsuki

    Genes to Cells   23 ( 3 )   225 - 233   2018年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Blackwell Publishing Ltd  

    MicroRNAs (miRNAs) belong to a class of small noncoding RNAs that play important roles in the translational regulation of gene expression. A number of miRNAs are known to act as key regulators of diverse processes such as neuronal differentiation. In this study, we have attempted to identify novel miRNAs related to neuronal differentiation via microarray analysis in the human neuronal differentiation model neuroblastoma SH-SY5Y cells. We identified 15 up-regulated and eight down-regulated miRNAs in SH-SY5Y cells treated with all-trans retinoic acid to induce differentiation. We further showed that one of the up-regulated miRNAs, miR-664a-5p, promoted neuronal differentiation of SH-SY5Y cells. These findings enhance our understanding of the miRNAs involved in the process of neurogenesis and, in particular, highlight an important role of miR-664a-5p in SH-SY5Y cell neuronal differentiation. Further studies will be required to confirm the function of miR-664-5p in neuronal development and disease and to identify its relevant target genes.

    DOI: 10.1111/gtc.12559

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

    Hyungjin Kim, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   28 ( 3 )   378 - 381   2018年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier Ltd  

    Multivalent cell-penetrating peptides (CPPs) have been reported to show enhancement in cellular uptake and endosomolytic activity. However, its application was limited to trans-delivery of cargo which is lower in cellular uptake efficiency of cargo than cis-delivery. Here, we tried the cis-delivery of cargo with multivalent CPP by preparing bioreducible dimeric CPP–cargo with apoptotic activity using TatBim peptide, a fusion of Tat CPP and Bim peptide derived from Bim apoptosis-inducing protein. Dimeric TatBim was almost twice as highly internalized by cells and significantly induced apoptosis compared to monomeric TatBim. Contribution of bioreducible linkage of dimeric TatBim towards apoptotic activity was also confirmed.

    DOI: 10.1016/j.bmcl.2017.12.035

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  • Ultrasound-dependent cytoplasmic internalization of a peptidesonosensitizer conjugate 査読

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

    Bioorganic & Medicinal Chemistry   25 ( 15 )   4212 - 4217   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    A method to induce cytoplasmic peptide delivery, using ultrasound, was demonstrated using a molecular conjugate of a cell-penetrating peptide (CPP), a functional peptide, and a sonosensitizer. As a model of such molecular conjugates, TatBim-RB, consisting of the Tat CPP, the Bim apoptosis inducing peptide, and the sonosensitizer rose bengal was synthesized. CPPs have been widely used for intracellular delivery of various cargos; however, CPP-fused molecules tend to become entrapped in endosomes, as was observed for TatBim-RB molecules applied to cells. To promote escape of the entrapped TatBim-RB molecules, cells were irradiated with ultrasound, which successfully induced endosomal escape and cytoplasmic dispersion of TatBim-RB, and subsequently apoptosis. Our results suggest that this peptidesonosensitizer conjugate strategy may facilitate numerous kinds of medicinal chemistry studies, and furthermore, this specific conjugate may exhibit potential as a novel therapeutic agent for the promotion of apoptosis. (C) 2017 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.bmc.2017.06.024

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

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

    Analytical Methods   9 ( 20 )   2971 - 2976   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROYAL SOC CHEMISTRY  

    The use of molecular beacons is a versatile method to detect RNAs. Typically, a single-stranded region of RNA is selected as a target sequence for molecular beacons. Therefore, the detection of highly structured short RNAs, such as tRNAs, seems to be difficult. In this study, as an example of highly structured target RNA, we used human tRNA(Lys3), which is known to have functions in protein synthesis and the reverse transcription of human immunodeficiency virus (HIV)-1. No long single-stranded region more than 8 nt is present in this tRNA, which is much shorter than the standard target length of molecular beacons (similar to 20 nt). This study showed that sensitive detection of highly structured RNAs using molecular beacons was much more difficult than that of unstructured or less structured RNAs. However, efficient detection of the tRNA(Lys3) was achieved by selecting the best target region, i.e., the region around the D arm, probably due to the ease of unfolding of this arm. Accordingly, our findings suggested that molecular beacons may have applications in the detection of highly structured RNAs related to various biological functions and diseases.

    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 査読

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

    Biochemical Journal   474   957 - 969   2017年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PORTLAND PRESS LTD  

    Translation elongation factor Tu (EF-Tu) delivers aminoacyl-tRNA (aa-tRNA) to ribosomes in protein synthesis. EF-Tu generally recognizes aminoacyl moieties and acceptor-and T-stems of aa-tRNAs. However, nematode mitochondrial (mt) tRNAs frequently lack all or part of the T-arm that is recognized by canonical EF-Tu. We previously reported that two distinct EF-Tu species, EF-Tu1 and EF-Tu2, respectively, recognize mt tRNAs lacking T-arms and D-arms in the mitochondria of the chromadorean nematode Caenorhabditis elegans. C. elegans EF-Tu2 specifically recognizes the seryl moiety of serylated D-armless tRNAs. Mitochondria of the enoplean nematode Trichinella possess three structural types of tRNAs: T-armless tRNAs, D-armless tRNAs, and cloverleaf tRNAs with a short T-arm. Trichinella mt EF-Tu1 binds to all three types and EF-Tu2 binds only to D-armless Ser-tRNAs, showing an evolutionary intermediate state from canonical EF-Tu to chromadorean nematode (e. g. C. elegans) EF-Tu species. We report here that two EF-Tu species also participate in Drosophila melanogaster mitochondria. Both D. melanogaster EF-Tu1 and EF-Tu2 bound to cloverleaf and D-armless tRNAs. D. melanogaster EF-Tu1 has the ability to recognize T-armless tRNAs that do not evidently exist in D. melanogaster mitochondria, but do exist in related arthropod species. In addition, D. melanogaster EF-Tu2 preferentially bound to aatRNAs carrying small amino acids, but not to aa-tRNAs carrying bulky amino acids. These results suggest that the Drosophila mt translation system could be another intermediate state between the canonical and nematode mitochondria-type translation systems.

    DOI: 10.1042/BCJ20160929

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

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

    Nature Communications   7   12501 - 12501   2016年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl) methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4 h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at similar to 405 nm at 125mWcm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19 s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells.

    DOI: 10.1038/ncomms12501

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  • Photoinduced apoptosis using a peptide carrying a photosensitizer 査読

    Kazunori Watanabe, Hayato Fujiwara, Mizuki Kitamatsu, Takashi Ohtsuki

    Bioorganic & Medicinal Chemistry Letters   26 ( 13 )   3115 - 3118   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    A novel molecule, TatBim-Alexa, consisting of the HIV1 Tat cell-penetrating peptide, the Bim apoptosis-inducing peptide, and Alexa Fluor 546 was synthesized for photoinducion of apoptosis. The Alexa Fluor 546 was used as a photosensitizer and covalently attached at the C-terminus of TatBim peptide by the thiol-maleimide reaction. Photo-dependent cytosolic internalization of TatBim-Alexa and photo-dependent apoptosis using TatBim-Alexa were demonstrated in several kinds of mammalian cells including human cancer cell lines. (C) 2016 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.bmcl.2016.04.091

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  • Enhanced cellular uptake of lactosomes using cell-penetrating peptides. 査読

    Akahoshi, A, Matuura, E, Ozeki, E, Matsui, H, Watanabe, K, Ohtsuki, T

    Sci. Technol. Adv. Mater.   17 ( 1 )   245 - 252   2016年6月

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  • Photocontrolled intracellular RNA delivery by using nanoparticles or carrier-photosensitizer conjugates 招待 査読

    Kazunori Watanabe, Takashi Ohtsuki

    Prog Mol Biol Transl Sci.   139   101 - 119   2016年

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    記述言語:英語  

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

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

    Scientific Reports   5   18577 - 18577   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    In many drug delivery strategies, an inefficient transfer of macromolecules such as proteins and nucleic acids to the cytosol often occurs because of their endosomal entrapment. One of the methods to overcome this problem is photochemical internalization, which is achieved using a photosensitizer and light to facilitate the endosomal escape of the macromolecule. In this study, we examined the molecular mechanism of photochemical internalization of cell penetrating peptide-cargo (macromolecule)-photosensitizer conjugates. We measured the photophysical properties of eight dyes (photosensitizer candidates) and determined the respective endosomal escape efficiencies using these dyes. Correlation plots between these factors indicated that the photogenerated O-1(2) molecules from photosensitizers were highly related to the endosomal escape efficiencies. The contribution of O-1(2) was confirmed using O-1(2) quenchers. In addition, time-lapse fluorescence imaging showed that the photoinduced endosomal escape occurred at a few seconds to a few minutes after irradiation (much longer than O-1(2) lifetime), and that the pH increased in the endosome prior to the endosomal escape of the macromolecule.

    DOI: 10.1038/srep18577

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

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

    Chemical Communications   51   413 - 416   2015年5月

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

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

    Bioorganic & Medicinal Chemistry Letters   24 ( 23 )   5369 - 5372   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Translation systems with four-base codons provide a powerful strategy for protein engineering and protein studies because they enable site-specific incorporation of non-natural amino acids into proteins. In this study, a caged aminoacyl-tRNA with a four-base anticodon was synthesized. The caged aminoacyl-tRNA contains a photocleavable nitroveratryloxycarbonyl (NVOC) group. This study showed that the caged aminoacyl-tRNA was not deacylated, did not bind to EF-Tu, and was activated by light. Photo-dependent translation of an mRNA containing the four-base codon was demonstrated using the caged aminoacyl-tRNA. (C) 2014 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.bmcl.2014.10.053

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

    Kazunori Watanabe, Kenichi Ijiri, Takashi Ohtsuki

    FEBS Letters   588 ( 18 )   3454 - 3460   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Stress induces various responses, including translational suppression and tRNA degradation in mammals. Previously, we showed that heat stress induces degradation of initiator tRNA(Met) (iMet) through 5'-3' exoribonuclease Xrn1 and Xrn2, respectively. In addition, we found that rapamycin inhibits the degradation of iMet under heat stress conditions. Here, we report that the mammalian target of rapamycin (mTOR) regulates the diffusion of Xrn2 from the nucleolus to the nucleoplasm, facilitating the degradation of iMet under conditions of heat stress. Our results suggest a mechanism of translational suppression through mTOR-regulated iMet degradation in mammalian cells. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    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 査読

    Yoh-ichi Watanabe, Takuma Suematsu, Takashi Ohtsuki

    Frontiers in Genetics   5   109   2014年5月

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    記述言語:英語   出版者・発行元:FRONTIERS MEDIA SA  

    Conventional tRNAs have highly conserved sequences, four-armed cloverleaf secondary structures, and L-shaped tertiary structures. However, metazoan mitochondrial tRNAs contain several exceptional structures. Almost all tRNA(Ser) for AGY/N codons lack the D-arm. Furthermore, in some nematodes, no four-armed cloverleaf-type tRNAs are present: two tRNAs(Ser) without the D-arm and 20 tRNAs without the T-arm are found. Previously, we showed that in nematode mitochondria, an extra elongation factor Tu (EF-Tu) has evolved to support interaction with tRNAs lacking the T-arm, which interact with C-terminal domain 3 in conventional EF-Tu. Recent mitochondrial genome analyses have suggested that in metazoan lineages other than nematodes, tRNAs without the T-arm are present. Furthermore, even more simplified tRNAs are predicted in some lineages. In this review, we discuss mitochondrial tRNAs with divergent structures, as well as protein factors, including EF-Tu, that support the function of truncated metazoan mitochondrial tRNAs.

    DOI: 10.3389/fgene.2014.00109

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  • Intracellular Delivery of RNA via RNA-Binding Proteins or Peptides 招待 査読

    Kazunori Watanabe, Takashi Ohtsuki

    Intracellular Delivery II   403 - 416   2014年

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    掲載種別:論文集(書籍)内論文   出版者・発行元:Springer Netherlands  

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

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

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

    Bioconjugate Chemistry   24 ( 10 )   1669 - 1673   2013年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Controlled activation of small RNAs, such as small interfering RNA, in cells is very useful for various biological applications. Light is an effective inducer of controlled activation
    in particular, near-infrared light is favorable because it can penetrate deeper into tissues than UV or visible light. In this study, near-infrared light control of RNA interference (RNAi) was demonstrated in mammalian cells using a photosensitive RNA carrier molecule, consisting of an RNA carrier protein and a fluorochrome. The photosensitive carrier molecule was identified from six candidates, each with a different fluorochrome. Using this carrier molecule, cytosolic RNA delivery and RNAi can be triggered by near-infrared light. Cytotoxicity was not observed after photoinduction of RNAi. © 2013 American Chemical Society.

    DOI: 10.1021/bc4001195

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

    Mizuki Kitamatsu, Mayumi Kitabatake, Yoshiteru Noutoshi, Takashi Ohtsuki

    Biopolymers   100 ( 1 )   64 - 70   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In this report, we describe dendritic peptides possessing central fluorescent amino acids with adjacent branched amino acids. These fluorescent-peptide dendrimers were prepared using (9-fluorenyl)methoxycarbonyl (Fmoc)-based solid-phase peptide synthesis and Fmoc-derivative fluorescent and branched amino acids. The branched amino acids featured multiple carboxylic acids in their side chains, making the fluorescent-peptide dendrimers highly water-soluble compared with the analogous peptides containing the fluorescent amino acids only. The branched amino acid units also improved the fluorescence intensity of the dendrimers. Based on high-pressure liquid chromatography and fluorescence spectroscopy results, we determined that the fluorescent groups were located in the core and that the carboxylic acids were located on the surface of the dendrimers. Fluorescence resonance energy transfer was achieved among the three proximal fluorescent groups in one of the fabricated fluorescent-peptide dendrimers. (C) 2012 Wiley Periodicals, Inc.

    DOI: 10.1002/bip.22175

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  • Photoinduced RNA Interference 招待 査読

    Yuka Matsushita-Ishiodori, Takashi Ohtsuki

    Accounts of Chemical Research   45 ( 7 )   1039 - 1047   2012年7月

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    記述言語:英語   出版者・発行元:AMER CHEMICAL SOC  

    Because RNA interference (RNAi) can be applied to any gene, this technique has been widely used for studying gene functions. In addition, many researchers are attempting to use RNAi technology in RNAi-based therapies. However, several challenging and controversial issues have arisen during the widespread application of RNAi including target gene specificity, target cell specificity, and spatiotemporal control of gene silencing. To address these issues, several groups have utilized photochemistry to control the RNA release, both spatially and temporally.
    In this Account, we foam on recent studies using photocleavable protecting groups, photosensitizes, Hand gold nanoparticles for photoinduced RNAi. In 2005 the first report of photoinduced RNAi used a caged short interfering RNA (siRNA), an siRNA carrying a photocleavable protecting group. Caging groups block the bioactivities of target molecules, but allow for complete recovery of these functions via photoactivation. However, some RNAi activity can occur in these caged siRNAs, so it will be necessary to decrease this "leakage" and raise the RNAi activity restored after irradiation. This technique also uses UV light around 350 nm, which is cytotoxic, but in the near future we expect that it will be possible to use visible and near-infrared light
    We also examine the application of photochemical internalization (PCI) to RNAi technology, which involves a combination of photosensitizers and light Instead of inducing RNAi using light, the strategy behind this method was to enhance RNAi using RNA carriers. Many wellknown RNA carriers deliver siRNAs into cells by endocytosis. The siRNAs are trapped in endocytic vesicles and have to be released into the cytoplasm in order to express their activity. To achieve the endosomal escape of siRNAs, PCI technology employed photosensitizers to generate light-dependent reactive oxygen species (ROS) that disrupted the endocytic vesicles. In most studies, RNAi-mediated knockdown of the target gene was detected even without PCI. Recently, a polymer capable of trapping the siRNA in endocytic vesicles controlled RNAi almost entirely by light. CLIP-RNAi uses photosensitizing carrier proteins that can be activated over a wide range of visible light wavelengths. With this method RNA carrier/siRNA complexes are completely trapped within endosomes, and RNAi is controlled strictly by light. Such precise, light-dependent control will open up new possibilities for cellular and molecular biology and therapy.
    Most recently, gold nanoparticles (AuNPs) conjugated to siRNA have provided temporal and spatial control of RNAL The light-dependent melting of AuNPs accompanied by a shape transformation induces the release of thiolated siRNAs from AuNPs. In this method, the unique optical properties of the AuNP enable deep penetration of the excitation light into tissues at nearinfrared wavelengths.
    The development of photoinduced RNAi technology will lead to novel insights into gene functions and selective drug delivery, and many other scientific fields will continue to influence its progress.

    DOI: 10.1021/ar200227n

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  • Photosensitizing Carrier Proteins for Photoinducible RNA Interference 査読

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

    Bioconjugate Chemistry   22 ( 11 )   2222 - 2226   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    RNA interference (RNAi) is being widely explored as a tool in functional genomics and tissue engineering, and in the therapy of intractable diseases, including cancer and neurodegenerative diseases. Recently, we developed a photoinducible RNAi method using photosensitizing carrier proteins, named CLIP-RNAi (CPP-linked REP-mediated RNA internalization and photoinduced RNAi). Novel carrier proteins were designed for this study to establish a highly efficient delivery system for small interfering RNA (siRNA) or short hairpin RNA (shRNA) and to demonstrate light-dependent gene silencing. In addition, the results suggested that the dissociation of the siRNA (or shRNA) from carrier proteins in the cytoplasm is a critical event in CLIP-RNAi-mediated gene silencing.

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

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

    Analytical Biochemistry   418 ( 1 )   97 - 101   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The integral membrane protein bacteriorhodopsin, containing a fluorescent amino acid at a specific position, was synthesized in the presence of hydrated lipid films using an in vitro translation system expanded with a four-base codon/anticodon pair. Cell-sized liposomes with the labeled protein inserted into the liposome membranes were generated after the translation reaction. This study also demonstrated that this labeling method could be used to analyze the dynamic properties of membrane proteins in situ by fluorescence correlation spectroscopy. (C) 2011 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.ab.2011.06.026

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

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

    Biochem. Biophys. Res. Commun.   414 ( 3 )   625 - 630   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Arginine analogs were incorporated site-specifically into proteins using an in vitro translation system. In this system, mRNAs containing a CGGG codon were translated by an aminoacyl-tRNA(CCCG), which was charged with arginine analogs using yeast arginyl-tRNA synthetase. N(G)-monomethyl-L-arginine, L-citrulline and L-homoarginine were incorporated successfully into proteins using this method. The influence of arginine monomethylation in histone H3 on the acetylation of lysine residues by histone acetyltransferase hGCN5 was investigated, and the results demonstrated that K9 acetylation was suppressed by the methylation of R8 and R17 but not by R26 methylation. K18 acetylation was not affected by the methylation of R8. R17 and R26. This site-specific modification strategy provides a way to explore the roles of post-translational modifications in the absence of heterogeneity due to other modifications. (C) 2011 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2011.09.137

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

    Mizuki Kitamatsu, Shunsuke Kurami, Takashi Ohtsuki, Masahiko Sisido

    Bioorganic & Medicinal Chemistry Letters   21 ( 1 )   225 - 227   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    To investigate the antisense effect of a pyrrolidine-based oxy-peptide nucleic acid (POPNA), we carried out the LacZ reporter assay using a 12-mer trans-L-POPNA conjugated with a cell-penetrating peptide (antisense reagent). The antisense effect of the conjugated POPNA (inhibition of LacZ activity) was comparable to that shown by a Nielsen-type peptide nucleic acid. Furthermore, the conjugated POPNA could switch the LacZ activity over a wide range of ambient temperatures. (C) 2010 Elsevier Ltd. All rights reserved.

    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 査読

    Toshimasa Hamamoto, Masahiko Sisido, Takashi Ohtsuki, Masumi Taki

    Chemical Communications   47 ( 32 )   9116 - 9118   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROYAL SOC CHEMISTRY  

    By using the NEXT-A reaction, we introduced a non-natural amino acid at the N-terminus of a peptide/protein that contained a cysteine unit. The side chain of the introduced amino acid spontaneously reacted with the cysteine to afford a cyclic peptide/protein.

    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 査読

    Mizuki Kitamatsu, Akiko Takahashi, Takashi Ohtsuki, Masahiko Sisido

    Tetrahedron   66 ( 51 )   9659 - 9666   2010年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    We synthesized 16 pyrrolidine-based oxy-peptide nucleic acid (POPNA) monomers carrying four different nucleobases onto four different stereoisomers of pyrrolidine rings. Using these monomers, we prepared POPNA oligomers, which formed sequence-specific hybrids with DNAs. The oligomer configurations influenced the hybrid stability. The oligomers were not taken into CHO cells. However, they could enter the cell cytoplasm when mixed with the influenza virus hemagglutinin peptide-arginine heptamer conjugate. (C) 2010 Elsevier Ltd. All rights reserved.

    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 査読

    Takashi Ohtsuki, Hiromichi Yamamoto, Yoshio Doi, Masahiko Sisido

    Journal of Biochemistry   148 ( 2 )   239 - 246   2010年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    We present three methods relating to tRNA aminoacylation with non-natural amino acids using an Escherichia coli EF-Tu E215A/D216A mutant that can bind tightly to aa-tRNAs carrying either non-natural or natural amino acids: (i) a method for improving aminoacylation efficiency, (ii) a rapid method for analysing aminoacylation efficiency without the use of radioisotope labelling and (iii) a method for purifying aminoacyl-tRNAs. Although the EF-Tu mutant may be incompatible with some kinds of non-natural amino acids, we confirmed that the EF-Tu mutant could efficiently bind to aa-tRNAs carrying various amino acids (Arg, Ser, O-methyltyrosine, Bodipy FL-aminophenylalanine and 2-acrydonylalanine). These methods may be used for the efficient in vitro synthesis of proteins containing various non-natural amino acids.

    DOI: 10.1093/jb/mvq053

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  • Lactobacillus-mediated RNA interference in nematode 査読

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

    J. Biosci. Bioeng.   109 ( 2 )   189 - 192   2010年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    We engineered Lactobacillus paracasei to produce a dsRNA that would trigger RNAi-induced silencing of an essential gene in the nematode Caenorhabditis elegans. The dsRNA-expressing L paracasei can be used in experiments conducted on culture plates and may also be used as an orally administrable dsRNA carrier for humans and other mammals. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.

    DOI: 10.1016/j.jbiosc.2009.08.002

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  • Cellular siRNA Delivery Using TatU1A and Photo-Induced RNA Interference 招待 査読

    Tamaki Endoh, Takashi Ohtsuki

    RNA Interference   271 - 281   2010年

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    掲載種別:論文集(書籍)内論文   出版者・発行元:Humana Press  

    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 査読

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

    Bioconjugate Chemistry   20 ( 12 )   2242 - 2246   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    Bioactive small molecules such as metabolites and drugs play important roles in regulating biological functions. A technique for Visualizing such small molecules is very useful to understand their molecular mechanisms. In this study, an RNA-protein Conjugate, which consists of in RRE-RNA sensor protein (EYFP-Rev-ECFP) and an altered RRE-RNA, was constructed to detect bioactive small molecules by fluorescent resonance energy transfer (FRET). We designed a theophylline-aptamer-inserted RRE-RNA (Theo-RRE) to detect theophylline as a model target molecule. Theo-RRE formed an RNA-protein conjugate with EYFP-Rev-ECFP in the presence of theophylline. As a result, theophylline was specifically detected down to 10 mu M by the FRET increase in distinction from theophylline analogue, caffeine, in cell lysates.

    DOI: 10.1021/bc9002184

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

    Tamaki Endoh, Masahiko Sisido, Takashi Ohtsuki

    Journal of Controlled Release   137 ( 3-4 )   241 - 245   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    In this study we describe the spatial regulation of RNA interference (RNAi) using an RNA-carrier protein labeled with a fluorescent dye and a light source to trigger the RNAi. We demonstrate photo-dependent gene silencing using several dyes with different excitation wavelengths. Additionally, we use light from a halogen lamp and a photomask to produce photopatterned RNAi, and laser light to trigger single-cell RNAi on cell culture plates. (C) 2009 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jconrel.2009.04.015

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  • Cellular siRNA delivery using cell-penetrating peptides modified for endosomal escape 招待 査読

    Tamaki Endoh, Takashi Ohtsuki

    Advanced Drug Delivery Reviews   61 ( 9 )   704 - 709   2009年7月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    RNAi-mediated silencing of specific genes is a promising strategy for gene therapy. To utilize RNAi for therapy, an efficient and safe method for delivery of RNA into the cell cytosol is necessary. The plasma membrane is the primary, and most difficult, barrier for RNA to cross, because negatively charged RNA is strongly repulsed by the negatively charged membrane. A variety of cationic polymers can be used as RNA carriers by interacting with RNA and covering its negative charges to form a cell-penetrating complex Among the emerging candidates for RNA carriers are cationic cell-penetrating peptides (CPPs), which can cross the plasma membrane and internalize into cells together with RNA. This review focuses on CPP-based RNA delivery strategies. In using CPP-based RNA delivery, most of the RNA internalized by the cell is entrapped in endosomes. Strategies for endosomal escape of RNAs are also reviewed. (C) 2009 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.addr.2009.04.005

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  • Carrier PNA for shRNA delivery into cells 査読

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

    Bioorganic & Medicinal Chemistry Letters   19 ( 13 )   3410 - 3413   2009年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    A peptide nucleic acid (PNA)-cell-penetrating peptide (CPP) conjugate (carrier PNA) was used as 'bridgebuilder' to connect a CPP with an shRNA. The carrier PNA successfully formed a hybrid with an shRNA bearing complementary dangling bases and the shRNA was introduced into cells by the carrier PNA, and RNAi was induced by the shRNA. Crown Copyright (C) 2009 Published by Elsevier Ltd. All rights reserved.

    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 査読

    Hidetaka Nakata, Takashi Ohtsuki, Masahiko Sisido

    Analytical Biochemistry   390 ( 2 )   121 - 125   2009年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We developed novel Substrates for protease activity evaluation by fluorescence correlation spectroscopy (FCS). Substrates were labeled in a position-specific manner with a fluorophore near the N terminus and included a C-terminal, 30 kDa, highly soluble protein (elongation factor Ts [EF-Ts]). The C-terminal protein enhanced the substrate peptide Solubility and increased the molecular weight, enabling sensitive detection by FCS. Using the labeled substrates, caspase-3 and matrix metalloproteinase-9 (MMP-9) activities were confirmed by FCS. To demonstrate the Suitability of this FCS-based assay for high-throughput screening, we screened various chemical compounds for MMP-9 inhibitors. The screening results confirmed the inhibitory activity of one compound and also revealed another potential MMP-9 inhibitor. Thus, this combination of position-specific labeled protein Substrates and FCS may serve as a useful tool for evaluating activities of various proteases and for protease inhibitor screening. (C) 2009 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.ab.2009.03.049

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  • PNA Arrays for miRNA Detection 査読

    Tamaki Endoh, Mizuki Kitamatsu, Masahiko Sisido, Takashi Ohtsuki

    Chemistry Letters   38 ( 5 )   438 - 439   2009年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHEMICAL SOC JAPAN  

    The profiling of microRNAs (miRNAs) using microarray technology has been used in many recent biological and medical studies. In this study, arrays of peptide nucleic acids (PNAs) were prepared and their miRNA capture abilities were evaluated. We found that the sensitivity of 10-mer PNA probes in targeting miRNAs was much higher than that of the corresponding DNA probes and also of longer (20-mer) DNA probes.

    DOI: 10.1246/cl.2009.438

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  • The Central Dogma: From DNA to RNA, and to Protein 査読

    Takashi Ohtsuki, Masahiko Sisido

    Automation in Proteomics and Genomics   1 - 19   2009年3月

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    記述言語:英語   掲載種別:論文集(書籍)内論文   出版者・発行元:John Wiley & Sons, Ltd  

    DOI: 10.1002/9780470741191.ch1

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

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

    Journal of Organic Chemistry   74 ( 6 )   2585 - 2588   2009年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    Unique taurine-containing Uridine derivatives, 5-taurinomethyluridine (tau m(5)U) and 5-taurinomethyl-2-thiouridine (tau m(5)s(2)U), which were discovered in mammalian mitochondrial tRNAs, exist at the first position of the anticodon. In this paper, we report the first efficient synthesis of tau m(5)U and tau m(5)s(2)U and describe their physicochemical properties. These modified ribonucleosides were synthesized by the reaction of 5-substituted uridine derivatives with a tetrabutylammonium salt of taurine that is highly reactive and well-soluble in common organic solvents. UV and (1)H NMR spectrometric studies revealed the structural properties of the taurine-containing base moieties and the Sugar conformations of these modified ribonucleosides.

    DOI: 10.1021/jo802697r

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  • Isolation of small RNAs using biotinylated PNAs

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

    Journal of Biochemistry   144 ( 4 )   415 - 418   2008年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    In this study, an RNA isolation method was developed using a biotinylated peptide nucleic acid (PNA) that is complementary to the target RNA. Using the biotinylated PNA method, we successfully isolated several RNAs from Escherichia coli and from human total RNA in pure form. Damage to the RNA appears to be negligible by this method because the method is rapid and does not require a high temperature treatment to facilitate RNA-PNA binding.

    DOI: 10.1093/jb/mvn107

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  • Amino acid sensing using aminoacyl-tRNA synthetase

    Akimitsu Kugimiya, Miki Morii, Takashi Ohtsuki

    Analytical Biochemistry   378 ( 1 )   90 - 92   2008年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The detection of amino acids using aminoacyl-tRNA synthetases (ARSs) as the molecular recognition element was proposed, and the binding activity and specificity of ARSs were evaluated. Using this rapid and easy method, from 15 to 50 mu M tyrosine could be measured specifically. The method suggested in this article could be realized without an amino acid labeling process or a large volume of organic solvents, and the time for measurement was reasonable. (c) 2008 Elsevier Inc. All rights reserved.

    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

    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年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Post-transcriptional modifications at the first ( wobble) position of the tRNA anticodon participate in precise decoding of the genetic code. To decode codons that end in a purine (R) (i.e. NNR), tRNAs frequently utilize 5-methyluridine derivatives (xm(5)U) at the wobble position. However, the functional properties of the C5-substituents of xm(5)U in codon recognition remain elusive. We previously found that mitochondrial tRNAs(Leu(UUR)) with pathogenic point mutations isolated from MELAS ( mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) patients lacked the 5-taurinomethyluridine (tau m(5)U) modification and caused a decoding defect. Here, we constructed Escherichia coli tRNAs(Leu(UUR)) with or without xm(5)U modifications at the wobble position and measured their decoding activities in an in vitro translation as well as by A-site tRNA binding. In addition, the decoding properties of tRNA(Arg) lacking mnm(5)U modification in a knock-out strain of the modifying enzyme (Delta mnmE) were examined by pulse labeling using reporter constructs with consecutive AGR codons. Our results demonstrate that the xm(5)U modification plays a critical role in decoding NNG codons by stabilizing U center dot G pairing at the wobble position. Crystal structures of an anticodon stem-loop containing tau m(5)U interacting with a UUA or UUG codon at the ribosomal A-site revealed that the m(5)U center dot G base pair does not have classical U center dot G wobble geometry. These structures provide help to explain how the tau m(5)U modification enables efficient decoding of UUG codons.

    DOI: 10.1074/jbc.M800233200

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

    Tamaki Endoh, Masahiko Sisido, Takashi Ohtsuki

    Bioconjugate Chemistry   19 ( 5 )   1017 - 1024   2008年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    HIV-1 TAT peptide, which is a cell-penetrating peptide (CPP), was fused to the UIA RNA-binding domain (TatU1A) to generate a sequence-specific siRNA delivery system for mammalian cells. The siRNA contained a short 5'-extension that is specifically recognized by the U1A RNA-binding domain (U1AsiRNA). Specific binding of TatU1A to the U1AsiRNA was confirmed using a gel mobility shift assay. The U1AsiRNA was internalized by cells only when it was preincubated with TatU1A before addition to the cells. Although most of the internalized siRNA seemed. to be entrapped in endocytic compartments, efficient redistribution of the entrapped siRNAs was achieved by photostimulation of a fluorophore attached to TatU1A. Once in the cytoplasm, the siRNA induced RNAi-mediated gene silencing. We refer to this delivery strategy as CLIP-RNAi. CLIP-RNAi is a promising strategy for RNAi experiments and for pinpoint RNAi therapy.

    DOI: 10.1021/bc800020n

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

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

    J. Am. Chem. Soc.   129 ( 46 )   14458 - 14462   2007年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    Nonnatural amino acids have been introduced into proteins using expanded protein biosynthesis systems. However, some nonnatural amino acids, especially those containing large aromatic groups, are not efficiently incorporated into proteins. Reduced binding efficiency of aminoacylated tRNAs to elongation factor Tu (EF-Tu) is likely to limit incorporation of large amino acids. Our previous studies suggested that tRNAs carrying large nonnatural amino acids are bound less tightly to EF-Tu than natural amino acids. To expand the availability of nonnatural mutagenesis, EF-Tu from the E coli translation system was improved to accept such large amino acids. We synthesized EF-Tu mutants, in which the binding pocket of the aminoacyl moiety of aminoacyl-tRNA was enlarged. L-1-Pyrenylalanine, L-2-pyrenylalanine, and DL-2anthraquinonylalanine, which are hardly or only slightly incorporated with the wild-type EF-Tu, were successfully incorporated into a protein using these EF-Tu mutants.

    DOI: 10.1021/ja075557u

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  • T-armless tRNAs and elongated elongation factor Tu.

    Ohtsuki, T, Watanabe, Y

    IUBMB life   59 ( 2 )   68 - 75   2007年

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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

    Masumi Taki, Yasunori Tokuda, Takashi Ohtsuki, Masahiko Sisido

    J. Biosci. Bioeng.   102 ( 6 )   511 - 517   2006年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    Spodoptera frugiperda 21 (Sf21) insect cell-free protein synthesizing system was expanded to include nonnatural amino acids. Orthogonal tRNAs that work as carriers of nonnatural amino acids in the insect system were explored. Four-base codons for assigning the positions of nonnatural amino acids were also selected. Mutated streptavidin mRNAs that contained different four-base codons were prepared and added to the insect cell-free system in the presence of various tRNAs possessing the corresponding four-base anticodons. The tRNAs were chemically amino-acylated with various types of nonnatural amino acids to examine their incorporation efficiencies. Using p-nitrophenylalanine as the nonnatural amino acid and streptavidin as the target protein, tRNA sequences and the types of four-base codons were optimized to maximize the yield of the nonnatural mutant and to minimize production of full-length proteins that do not contain the nonnatural amino acid. Among the tRNA sequences taken from a variety of tRNAs of nonstandard structures, the tRNA derived from Methanosarcina acetivorans tRNA(Pyl) was the most efficient and orthogonal tRNA. Of the CGGN-type four-base codons, CGGA and CGGG were the most efficient ones for assigning the positions of nonnatural amino acids. p-Nitrophenylalanine and 2-naphthylalanine were efficiently incorporated as in the case of Escherichia coli and rabbit reticulocyte cell-free systems. Much less efficient incorporation was observed, however, for other nonnatural amino acids, indicating that the insect system is less tolerant to the structural diversity of amino acids than the E. coli cell-free system.

    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.

    Arita, M, Suematsu, T, Osanai, A, Inaba, T, Kamiya, H, Kita, K, Sisido, M, Watanabe, Y, Ohtsuki, T

    Nucleic Acids Research   34 ( 18 )   5291 - 5299   2006年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1093/nar/gkl526

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

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

    Biochemistry   45 ( 36 )   10920 - 10927   2006年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    In canonical translation systems, the single elongation factor Tu (EF-Tu) recognizes all elongator tRNAs. However, in Caenorhabditis elegans mitochondria, two distinct EF-Tu species, EF-Tu1 and EF-Tu2, recognize 20 species of T armless tRNA and two species of D armless tRNA(Ser), respectively. We previously reported that C. elegans mitochondrial EF-Tu2 specifically recognizes the serine moiety of serylated-tRNA. In this study, to identify the critical residues for the serine specificity in EF-Tu2, several residues in the amino acid binding pocket of bacterial EF-Tu were systematically replaced with corresponding EF-Tu2 residues, and the mutants were analyzed for their specificity for esterified amino acids attached to tRNAs. In this way, we obtained a bacterial EF-Tu mutant that acquired serine specificity after the introduction of 10 EF-Tu2 residues into its amino acid binding pocket. C. elegans EF-Tu2 mutants lacking serine specificity were also created by replacing seven or eight residues with bacterial residues. Further stressing the importance of these residues, we found that they are almost conserved in EF-Tu2 sequences of closely related nematodes. Thus, these three approaches reveal the critical residues essential for the unique serine specificity of C. elegans mitochondrial EF-Tu2.

    DOI: 10.1021/bi060536i

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

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

    Analytical Biochemistry   348 ( 2 )   321 - 323   2006年1月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    DOI: 10.1016/j.ab.2005.08.008

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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.

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

    Biochemical Journal   2006年

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

    T Ohtsuki, T Manabe, M Sisido

    FEBS Letters   579 ( 30 )   6769 - 6774   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The ability to introduce non-natural amino acids into proteins opens up new vistas for the study of protein structure and function. This approach requires suppressor tRNAs that deliver the non-natural amino acid to a ribosome associated with an mRNA containing an expanded codon. The suppressor tRNAs must be absolutely protected from aminoacylation by any of the aminoacyl-tRNA synthetases in the protein synthesizing system, or a natural amino acid will be incorporated instead of the non-natural amino acid. Here, we found that some tRNAs with non-standard structures could work as efficient four-base suppressors fulfilling the above orthogonal conditions. Using these tRNAs, we successfully demonstrated incorporation of three different non-natural amino acids into a single protein. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    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

    M Sisido, K Ninomiya, T Ohtsuki, T Hohsaka

    Methods   36 ( 3 )   270 - 278   2005年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Techniques for position-specific incorporation of non-natural amino acids in an in vitro protein synthesizing system are described. First, a PNA-assisted non-enzymatic tRNA aminoacylation with a variety of natural and non-natural amino acids is described. With this technique, one can aminoacylate a specific tRNA simply by adding a preformed amino acid activated ester-PNA conjugate into an in vitro protein biosynthesizing system. Second, the genetic code is expanded by introducing 4-base codons that can be exclusively translated to non-natural amino acids. The most advantageous point of the 4-base codon strategy is to introduce multiple amino acids into specific positions in single proteins by using mutually orthogonal 4-base codons and orthogonal tRNAs. An easy and quick method for preparation of tRNAs possessing 4-base anticodons is also described. Combination of the non-enzymatic aminoacylation and the 4-base codon/anticodon strategy gives an easy and widely applicable technique for incorporating a variety of non-natural amino acids into proteins in vitro. (c) 2005 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.ymeth.2005.04.009

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

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

    Biochemistry   44 ( 25 )   9232 - 9237   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    In the present study, mitochondrial ribosomes of the nematode Ascaris suum were isolated and their physiochernical properties were compared to ribosomes of Escherichia coli. The sedimentation coefficient and buoyant density of A. suum mitochondrial ribosomes were determined. The sedimentation coefficient of the intact monosome was about 55 S. The buoyant density of formaldehyde-fixed ribosomes in cesium chloride was 1.40 g/cm(3), which suggests that the nematode mitoribosomes have a much higher protein composition than other mitoribosomes. The diffusion coefficients obtained from dynamic light scattering measurements were (1.48 +/- 0.04) x 10(-7) cm(2) s(-1) for 55 S mitoribosomes and (1.74 +/- 0.04) X 10(-7) cm(2) S-1 for the 70 S E. coli monosome. The diameter of mitoribosomes was measured by dynamic light-scattering analysis and electron microscopy. Though the nematode mitoribosome has a larger size than the bacterial ribosome, it does not differ significantly in size from mammalian mitoribosomes.

    DOI: 10.1021/bi047833c

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

    M Sakurai, T Ohtsuki, T Suzuki, K Watanabe

    FEBS Letters   579 ( 13 )   2767 - 2772   2005年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    To understand the decoding property of nematode mitochondrial tRNAs with unusual secondary structures, post-transcriptional modifications at wobble positions of Ascaris suum mitochondrial tRNAs corresponding to two-codon families ending with a purine were analyzed. 5-Carboxymethylaminomethyluridine (cmnm(5)U) was identified at the wobble positions of tRNA(Lys), tRNA(Glu) and tRNA(Gln), while 5-carboxymethylaminomethyl-2-thiouridine (cmnm(5)s(2)U) was present in tRNA(UAA)(Leu) and tRNA(Trp). In most bacterial and mitochondrial tRNAs, the 2-thiouridine derivative is present in tRNAs for Lys, Gin and Gln. These is no report that cmnm(5)s(2)U is used in tRNALeu and tRNA(Trp). The unusual usage of wobble modifications might assist decoding of nematode mitochondrial mRNAs. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.febslet.2005.04.009

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

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

    Nucleic Acids Research   33 ( 15 )   4683 - 4691   2005年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Nematode mitochondria expresses two types of extremely truncated tRNAs that are specifically recognized by two distinct elongation factor Tu (EF-Tu) species named EF-Tu1 and EF-Tu2. This is unlike the canonical EF-Tu molecule that participates in the standard protein biosynthesis systems, which basically recognizes all elongator tRNAs. EF-Tu2 specifically recognizes Ser-tRNA(Ser) that lacks a D arm but has a short T arm. Our previous study led us to speculate the lack of the D arm may be essential for the tRNA recognition of EF-Tu2. However, here, we showed that the EF-Tu2 can bind to D arm-bearing Ser-tRNAs, in which the D-T arm interaction was weakened by the mutations. The ethylnitrosourea-modification interference assay showed that EF-Tu2 is unique, in that it interacts with the phosphate groups on the T stem on the side that is opposite to where canonical EF-Tu binds. The hydrolysis protection assay using several EF-Tu2 mutants then strongly suggests that seven C-terminal amino acid residues of EF-Tu2 are essential for its aminoacyl-tRNA-binding activity. Our results indicate that the formation of the nematode mitochondrial (mt) EF-Tu2/GTP/aminoacyl-tRNA ternary complex is probably supported by a unique interaction between the C-terminal extension of EF-Tu2 and the tRNA.

    DOI: 10.1093/nar/gki784

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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

    M Sakurai, T Ohtsuki, K Watanabe

    Nucleic Acids Research   33 ( 5 )   1653 - 1661   2005年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    The mitochondria of the nematode Ascaris suum have tRNAs with unusual secondary structures that lack either the T-arm or D-arm found in most other organisms. Of the twenty-two tRNA species present in the mitochondria of A. suum, twenty lack the entire T-arm and two serine tRNAs lack the D-arm. To understand how such unusual tRNAs work in the nematode mitochondrial translation system, we analyzed post-transcriptional modifications of 11 mitochondrial tRNA species purified from A. suum, 10 of which lacked a T-arm and one of which lacked a D-arm. The most characteristic feature of nematode mitochondrial tRNAs lacking a T-arm was the presence of 1-methyladenosine at position 9 (m(1)A(9)). Synthesis of T-armless tRNAs with or without the modified nucleoside showed that T-armless tRNAs without the modification had much lower aminoacylation and EF-Tu-binding activities than native tRNAs. The addition of a single methyl group to A9 of these tRNAs was sufficient to restore nearly native levels of amino-acylation and EF-Tu-binding activity as well as tertiary structure, suggesting that m(1)A(9w) is a key residue for the activity of T-armless tRNAs. Thus, m(1)A(9) is indispensable for the structure and function of T-armless tRNAs of nematode mitochondrial origin.

    DOI: 10.1093/nar/gki309

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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.

    Suzuki, T, Ohtsuki, T, Watanabe, K

    Endocytobiosis Cell Research   2004年

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

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

    J. Biol. Chem.   278 ( 51 )   51968 - 51973   2003年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    The (6-4) photoproduct formed by ultraviolet light is known as an alkali-labile DNA lesion. Strand breaks occur at (6-4) photoproducts when UV-irradiated DNA is treated with hot alkali. We have analyzed the degradation reaction of this photoproduct under alkaline conditions using synthetic oligonucleotides. A tetramer, d(GT(6-4) TC), was prepared, and its degradation in 50 mM KOH at 60 degreesC was monitored by high performance liquid chromatography. A single peak with a UV absorption spectrum similar to that of the starting material was detected after the reaction, and this compound was regarded as an intermediate before the strand break. The formation of this intermediate was compared with intermediates from the degradation of other alkali-labile lesions such as the abasic site, thymine glycol, and 5,6-dihydrothymine. The results strongly suggested that the first step of the alkali degradation of the (6- 4) photoproduct was the hydrolysis between the N3 and C4 positions of the 5'-pyrimidine component. Analyses by NMR spectroscopy and mass spectrometry supported the chemical structure of this product. Assays of the complex formation with XPC . HR23B and the translesion synthesis by DNA polymerase eta revealed that the biochemical properties are indistinguishable between the intact and hydrolyzed photoproducts.

    DOI: 10.1074/jbc.M307186200

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  • Quick two-step RNA ligation employing periodate oxidation

    S Kurata, T Ohtsuki, T Suzuki, K Watanabe

    Nucleic Acids Research   31 ( 22 )   2003年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    The introduction of modified or labeled nucleotides into RNA is a powerful RNA engineering tool as it enables us to investigate how native RNA modifications affect RNA function and structure. It also helps in the structural analysis of RNA. A modified nucleotide can be introduced into a specific position of RNA by the method of two-step enzymatic ligation of RNA fragments. However, this method requires a complicated purification step between the two ligation steps that results in low yields of the ligation product. Here we have developed a new ligation technique employing periodate oxide that eliminates this purification step. This increases the total yield of the ligation product and makes it a faster procedure.

    DOI: 10.1093/nar/gng145

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

    T Ohtsuki, M Sakurai, A Sato, K Watanabe

    Nucleic Acids Research   30 ( 24 )   5444 - 5451   2002年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Caenorhabditis elegans mitochondria have two elongation factor (EF)-Tu species, denoted EF-Tu1 and EF-Tu2. Recombinant nematode EF-Ts purified from Escherichia coli bound both of these molecules and also stimulated the translational activity of EF-Tu, indicating that the nematode EF-Ts homolog is a functional EF-Ts protein of mitochondria. Complexes formed by the interaction of nematode EF-Ts with EF-Tu1 and EF-Tu2 could be detected by native gel electrophoresis and purified by gel filtration. Although the nematode mitochondrial (mt) EF-Tu molecules are extremely unstable and easily form aggregates, native gel electrophoresis and gel filtration analysis revealed that EF-Tu.EF-Ts complexes are significantly more soluble. This indicates that nematode EF-Ts can be used to stabilize homologous EF-Tu molecules for experimental purposes. The EF-Ts bound to two eubacterial EF-Tu species (E.coli and Thermus thermophilus). Although the EF-Ts did not bind to bovine mt EF-Tu, it could bind to a chimeric nematode-bovine EF-Tu molecule containing domains 1 and 2 from bovine mt EF-Tu. Thus, the nematode EF-Ts appears to have a broad specificity for EF-Tu molecules from different species.

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  • A unique serine-specific elongation factor Tu found in nematode mitochondria

    T Ohtsuki, A Sato, Y Watanabe, K Watanabe

    Nature Structural Biology   9 ( 9 )   669 - 673   2002年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    The translation elongation factor Tu (EF-Tu) delivers aminoacyl-tRNAs to ribosomes by recognizing the tRNA acceptor and T stems. However, the unusual truncation observed in some animal mitochondrial tRNAs seems to prevent recognition by a canonical EF-Tu. For instance, nematode mitochondria contain tRNAs lacking a T or D arm. We recently found an atypical EF-Tu (EF-Tu1) specific for nematode mitochondrial tRNAs that lack the T arm. We have now discovered a second factor, EF-Tu2, which binds only to tRNAs that lack a D arm. EF-Tu2 seems unique in its amino acid specificity because it recognizes the aminoacyl moiety of seryl-tRNAs and the tRNA structure itself. Such EF-Tu evolution might explain tRNA structural divergence in animal mitochondria.

    DOI: 10.1038/nsb826

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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

    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年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CAMBRIDGE UNIV PRESS  

    In the second step of the two consecutive transesterifications of the self-splicing reaction of the group I intron, the conserved guanosine at the 31 terminus of the intron (omegaG) binds to the guanosine-binding site (GBS) in the intron. In the present study, we designed a 22-nt model RNA (GBS/omegaG) including the GBS and omegaG from the Tetrahymena group I intron, and determined the solution structure by NMR methods. In this structure, omegaG is recognized by the formation of a base triple with the G2649.C311 base pair, and this recognition is stabilized by the stacking interaction between omegaG and C262. The bulged structure at A263 causes a large helical twist angle (40 +/- 8degrees) between the G264.C311 and C262.G312 base pairs. We named this type of binding pocket with a bulge and a large twist, formed on the major groove, a "Bulge-and-Twist" (BT) pocket. With another twist angle between the C262.G312 and G413.C313 base pairs (45 +/- 10degrees), the axis of GBS/omegaG is kinked at the GBS region. This kinked axis superimposes well on that of the corresponding region in the structure model built on a 5.0 Angstrom resolution electron density map (Golden et al., Science, 1998, 282:345-358). This compact structure of the GBS is also consistent with previous biochemical studies on group I introns. The BT pockets are also found in the arginine-binding site of the HIV-TAR RNA, and within the 16S rRNA and the 23S rRNA.

    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

    T Ohtsuki, G Kawai, K Watanabe

    FEBS Letters   514 ( 1 )   37 - 43   2002年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The tertiary structure of Ascaris suum mitochondrial tRNA(VCU)(Ser) was examined by nuclear magnetic resonance analysis using its transcript, since tRNA(UCU)(Ser), lacking the D arm and possessing a truncated T arm, is the shortest of all the known tRNAs. Most basepairs in the proposed secondary structure of tRNA(UCU)(Ser) were shown to exist, but the connector region comprising the truncated D loop and the extra loop was flexible. This flexibility, would enable adjustment of the mutual distance between the 3'-terminus and the anticodon consistent with that of usual tRNAs. Thus, tRNA(UCU)(Ser) appears to function in a similar way to that of usual tRNAs in the ribosome. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

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

    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年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE AMERICA INC  

    An unnatural base pair of 2-amino-6-(2-thienyl) purine (denoted by s) and pyridin-2-one (denoted by y) was developed to expand the genetic code. The ribonucleoside triphosphate of y was site-specifically incorporated into RNA, opposite s in a template, by T7 RNA polymerase. This transcription was coupled with translation in an Escherichia coli cell-free system. The yAG codon in the transcribed ras mRNA was recognized by the CUs anticodon of a yeast tyrosine transfer RNA (tRNA) variant, which had been enzymatically aminoacylated with an unnatural amino acid, 3-chlorotyrosine. Site-specific incorporation of 3-chlorotyrosine into the Ras protein was demonstrated by liquid chromatography-mass spectrometry (LC-MS) analysis of the products. This coupled transcription-translation system will permit the efficient synthesis of proteins with a tyrosine analog at the desired position.

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

    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年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    We have found the gene for a translation elongation factor Tu (EF-Tu) homologue in the genome of the nematode Caenorhabditis elegans, Because the corresponding protein was detected immunologically in a nematode mitochondrial (mt) extract, it could be regarded as a nematode mt EF-Tu. The protein possesses an extension of about 57 amino acids (we call this domain 3') at the C terminus, which is not found in any other known EF-Tu, Because most nematode mt tRNAs lack a T stem, domain 3' may be related to this feature. The nematode EF-Tu bound to nematode T stem-lacking tRNA, but bacterial EF-Tu was unable to do so. A series of domain exchange experiments strongly suggested that domains 3 and 3' are essential for binding to T stem-lacking tRNAs, This finding may constitute a novel example of the co-evolution of a structurally simplified RNA and the cognate RNA-binding protein, the latter having apparently acquired an additional domain to compensate for the lack of a binding site(s) on the RNA.

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  • Unnatural base pairs for specific transcription

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

    Proc. Natl. Acad. Sci. U.S.A.   98 ( 9 )   4922 - 4925   2001年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

    An unnatural base pair of 2-amino-6-(N,N-dimethylamino)purine (designated as x) and pyridin-2-one (designated as y) has been developed for specific transcription. The ribonucleoside triphosphates of y and a modified y, 5-methylpyridin-2-one, are selectively incorporated into RNA opposite x in the templates by T7 RNA polymerase, In addition, the sequences of the DNA templates containing x can be confirmed by a dideoxynucleotide chain-terminator method supplemented with the deoxynucleoside triphosphate of y, The bulky dimethylamino group of x in the templates effectively eliminates noncognate pairing with the natural bases. These results enable RNA biosynthesis for the specific incorporation of unnatural nucleotides at the desired positions.

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

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

    J. Am. Chem. Soc.   122 ( 25 )   6118 - 6119   2000年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

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

    T Ohtsuki, G Kawai, K Watanabe

    Journal of Biochemistry   124 ( 1 )   28 - 34   1998年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPANESE BIOCHEMICAL SOC  

    Most nematode mitochondrial (mt) tRNAs have a TV-replacement loop (TV loop) which replaces the normal T arm and the variable loop in standard tRNAs with a less-structured loop. The tertiary structure of such tRNAs has been discussed theoretically with reference to the crystal structure of yeast tRNA(Phe) [Wolstenholme et al, (1994) Nucleic Acids Res. 22, 4300-4306] and examined experimentally by chemical and enzymatic probing [Watanabe et al. (1994) J, Biol. Chem. 269, 22902-22906]. The results suggest that most regions of the tRNA other than the TV loop are folded in a similar manner to yeast tRNA(Phe). To confirm this notion more clearly, the tertiary structure of Ascaris suum mt tRNA(Met) was analyzed by NMR using various synthetic tRNAs site-specifically labeled with stable isotopes, which were prepared by a combination of chemical synthesis and enzymatic ligation, Tertiary interactions involving G(L2), G(L3), U(L4), and U8 were observed in the NMR spectra of the labeled tRNAs, but those relating to G(L5) were not. On the basis of these results, a possible tertiary structural model of nematode mitochondrial tRNA(Met) was constructed.

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

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

    Journal of Biochemistry   120 ( 6 )   1070 - 1073   1996年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JAPAN BIOCHEMICAL SOC  

    RNA samples corresponding to Ascaris suum mitochondrial tRNA(Met) were chemically and automatically synthesized in amounts sufficient for NMR measurement. Conventional and rapid deprotection methods gave tRNA samples with the same amino acid-accepting activity as those prepared by other method; enzymatic synthesis, and enzymatic ligation of chemically synthesized fragments. The synthetic tRNA showed the same H-1-NMR spectrum in the iminoproton region as the ligated tRNA, This rapid and reliable preparation method thus provides biologically active tRNA for NMR measurement, and further, it is applicable for synthesis of other large synthetic RNAs, by combining the site-specific isotopic labeling method.

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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

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

    Nucleic Acids Research   24 ( 4 )   662 - 667   1996年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS UNITED KINGDOM  

    Ascaris suum mitochondrial tRNA(Met) lacking the entire T stem was prepared by enzymatic ligation of two chemically synthesized RNA fragments. The synthetic tRNA could be charged with methionine by A.suum mitochondrial extract, although the charging activity was considerably low compared with that of the native tRNA, probably due to lack of modification, Enzymatic probing of the synthetic tRNA showed a very similar digestion pattern to that of the native tRNA(Met), which has already been concluded to take an L-shape-like structure [Watanabe et al, (1994) J. Biol. Chem., 269, 22902-22906], These results suggest that the synthetic tRNA possesses almost the same conformation as the native one, irrespective of the presence or absence of modified residues. The method of preparing the bizarre tRNA used here will provide a useful tool for elucidating the tertiary structure of such tRNAs, because they can be obtained without too much difficulty in the amounts necessary for physicochemical studies such as NMR spectroscopy.

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  • MOLLIFICATION OF CYTOTOXICITY OF SULFATED POLYSACCHARIDES BY FIBROBLAST GROWTH-FACTORS

    M KUNOU, T OHTSUKI, T AKAIKE, K HATANAKA

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

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MARCEL DEKKER INC  

    Sulfated polysaccharide which had a relatively high degree of sulfation showed cytotoxicity to 3T3-L1 fibroblasts. Acidic and basic fibroblast growth factors inhibited the cell damage caused by the sulfated polysaccharides, while epidermal growth factor and platelet growth factor had no effects.

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  • EFFECTS OF SYNTHETIC POLYANIONS ON 3T3-L1 FIBROBLAST PROLIFERATION STIMULATED BY FIBROBLAST GROWTH-FACTORS

    K HATANAKA, T OHTSUKI, M KUNOU

    Chemistry Letters   1407-1410 ( 8 )   1407 - 1410   1994年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHEMICAL SOC JAPAN  

    Effects of synthetic polyanions on the mitogenic activity of fibroblast growth factors were investigated. Poly(vinylsulfonate) contributed to potentiate the activity of the acidic fibroblast growth factor, while the carboxyl group was quite effective for basic fibroblast growth factor.

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書籍等出版物

  • 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年 

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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)( 担当: 共著)

    講談社サイエンティフク  2020年12月 

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    記述言語:日本語

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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)( 担当: 共著)

    化学同人  2020年4月 

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    記述言語:日本語

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

    木曽, 良明

    シーエムシー出版  2019年5月  ( ISBN:9784781314174

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    総ページ数:viii, 265p   記述言語:日本語

    CiNii Books

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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年 

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  • Intracellular Delivery II

    ( 担当: 共著)

    Springer  2014年  ( ISBN:9789401788953

  • Nanotechnology Tools for the Study of RNA(共著)

    Elsevier  2014年 

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  • RNA Interference

    Springer  2010年 

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  • 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年 

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

    Wiley  2009年 

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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年 

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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年 

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  • Cell-free translation system

    Springer-Verlag  2002年 

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  • 基礎生化学実験法 第4巻

    東京化学同人  2000年 

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MISC

  • 創薬を支える光技術

    渡邉 和則, 大槻 高史

    月刊 光アライアンス   ( 6月 )   1 - 4   2018年

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  • 89Zr標識ヒト抗体バリアントと新規DDSキャリアによるTheranostics技術

    竹中文章, 小林和子, 木村俊作, 小関英一, 大槻高史, 小渕浩嗣, 松浦栄次

    Drug Delivery System   33 ( 3 )   214 - 222   2018年

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  • 拡張翻訳系におけるケージド化合物の利用

    赤星彰也, 大槻高史

    化学   72   64 - 65   2017年

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  • 細胞内のタンパク質合成を、光でコントロールする!

    大槻高史

    Academist Journal   2016年

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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

    月刊 化学工業   66   34 - 39   2015年

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  • PCDR法とCLIP-RNAi法

    大槻高史

    生命化学研究レター   46   10 - 14   2014年

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  • 細胞内侵入性RNA結合蛋白質によるRNAデリバリー

    大槻高史

    高分子   58   140 - 141   2009年

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  • 人工核酸によるRNA検出

    大槻高史, 北松瑞生

    未来材料   9   16 - 21   2009年

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  • キャリアペプチドによるRNAの細胞内導入法

    大槻高史, 遠藤玉樹

    生化学   81 ( 2 )   110 - 112   2009年

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  • 改変EF-Tuを用いた翻訳系の拡張

    大槻高史

    日本化学会生体機能関連化学部会News Letter   22 ( 3 )   2 - 5   2007年

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  • 非天然アミノ酸の導入による蛋白質の蛍光ラベル法とその応用

    宍戸昌彦, 瀧真清, 大槻高史, 芳坂貴弘

    蛋白質核酸酵素   Vol.51, No.5 p399-407   2006年

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  • ミトコンドリアの翻訳システム

    渡辺公綱, 大槻高史, 鈴木勉

    蛋白質核酸酵素   48 ( 4 )   365 - 374   2003年

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講演・口頭発表等

  • 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年6月  日本DDS学会

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    開催年月日: 2017年6月

    記述言語:日本語  

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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年11月7日 

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    開催年月日: 2015年11月7日

    記述言語:日本語  

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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年 

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    開催年月日: 2014年

    記述言語:日本語  

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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年3月8日 

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    開催年月日: 2013年3月8日

    記述言語:日本語  

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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年3月8日 

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    開催年月日: 2013年3月8日

    記述言語:日本語  

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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年9月5日 

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    開催年月日: 2012年9月5日

    記述言語:日本語  

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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年11月7日 

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    開催年月日: 2011年11月7日

    記述言語:日本語  

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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年11月7日 

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    開催年月日: 2011年11月7日

    記述言語:日本語  

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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年9月1日 

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    開催年月日: 2011年9月1日

    記述言語:日本語  

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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年11月10日 

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    開催年月日: 2007年11月10日

    記述言語:日本語  

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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年7月23日 

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    開催年月日: 2007年7月23日

    記述言語:日本語  

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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年 

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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年 

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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年 

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  • CLIP-RNAi ~photoinduced RNAi strategy~.

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

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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年 

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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年 

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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年 

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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年 

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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年 

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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年 

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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年 

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

    JAACT2008  2008年 

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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年 

     詳細を見る

  • Improvement of CPP-RBD for siRNA delivery

    JAACT2008  2008年 

     詳細を見る

  • 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年 

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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年 

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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年 

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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

    第212回バイオロンジル会  2007年 

     詳細を見る

  • 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年 

     詳細を見る

  • RNA isolation using biotinylated PNAs

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

     詳細を見る

  • 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年 

     詳細を見る

  • 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

    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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

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

    12th annual meeting of RNA society  2007年 

     詳細を見る

  • RNA工学と拡張翻訳系

    第109回応用化学科セミナー  2007年 

     詳細を見る

  • 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年 

     詳細を見る

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

    12th annual meeting of RNA society  2007年 

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

    4th International Peptide Symposium  2007年 

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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年 

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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年 

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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年 

     詳細を見る

  • RNA interference using cell permeable protein carrier

    4th International Peptide Symposium  2007年 

     詳細を見る

  • 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年 

     詳細を見る

  • Photo inducible RNA interference using cell permeable protein carrier

    Endoh, T, Sisido, M, Ohtsuki, T

    5th International Symposium on Nucleic Acids Chemistry  2007年 

     詳細を見る

  • 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年 

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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年 

     詳細を見る

  • 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年 

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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

    第一回無細胞生命科学研究会  2006年 

     詳細を見る

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

    43JPS-PEM4  2006年 

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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年 

     詳細を見る

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

    NANOBIO-TOKYO2006  2006年 

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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年 

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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年 

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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年 

     詳細を見る

  • RNA isolation using biotinylated PNA.

    20th IUBMB International Congress of Biochemistry and Molecular Biology  2006年 

     詳細を見る

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

    20th IUBMB International Congress of Biochemistry and Molecular Biology  2006年 

     詳細を見る

  • 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年 

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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年 

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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年 

     詳細を見る

  • Specific RNA binding of immobilized PNAs

    29th European Peptide Symposium  2006年 

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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年 

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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年 

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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年 

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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

    企画セミナー「人工遺伝暗号」プログラム  2005年 

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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年 

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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年 

     詳細を見る

  • 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年 

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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年 

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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年 

     詳細を見る

  • 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年 

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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年 

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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年 

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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年 

     詳細を見る

  • 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年 

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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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

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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年 

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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年 

     詳細を見る

  • Elongation factors Tu for truncated tRNAs

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

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • Unique translation systems of animal mitochondria

    International congress on protein expression & protein function  2003年 

     詳細を見る

  • Modified nucleotides in nematode mitochondrial tRNAs

    20th International tRNA Workshop  2003年 

     詳細を見る

  • Unique serine-specificity of mitochondrial EF-Tu

    20th International tRNA Workshop  2003年 

     詳細を見る

  • 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年 

     詳細を見る

  • Unusual elongation factors Tu for truncated tRNAs

    20th International tRNA Workshop  2003年 

     詳細を見る

  • 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年 

     詳細を見る

  • Unique characteristics of animal mitochondrial translation systems

    20th International tRNA Workshop  2003年 

     詳細を見る

  • 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年 

     詳細を見る

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

    19th International tRNA workshop  2002年 

     詳細を見る

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

    19th International tRNA workshop  2002年 

     詳細を見る

  • 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年 

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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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

     詳細を見る

  • 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年 

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▼全件表示

産業財産権

  • 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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    出願番号:特願2019- 008817  出願日:2019年1月22日

    公開番号:特開2020-117479  公開日:2020年8月6日

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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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    出願人:株式会社島津製作所・岡山大学

    出願番号:特願2016-557827  出願日:2014年11月8日

    特許番号/登録番号:特許6586103  登録日:2019年9月13日 

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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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    出願番号:特願2011-063953  出願日:2011年11月24日

    特許番号/登録番号: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

    北松瑞生, 道上宏之, 王 飛霏, 中島真実, 大槻高史

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    出願番号:特願2011-233812  出願日:2011年10月25日

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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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    出願番号:特願2008-279228  出願日:2008年10月30日

    特許番号/登録番号: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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    出願番号:特願2008-1174884  出願日:2008年4月28日

    公開番号: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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    出願番号:特願2007-37041  出願日:2007年2月16日

    公開番号: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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    出願番号:特願2006-240828  出願日:2006年9月5日

    公開番号: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

     詳細を見る

    出願番号:特願2005-102868  出願日:2005年3月31日

    特許番号/登録番号: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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    出願番号:特願2005-103703  出願日:2005年3月31日

    特許番号/登録番号: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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    出願番号:特願2004-359328  出願日:2004年12月13日

    公開番号:特開2006-166709 

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▼全件表示

受賞

  • 研究功績賞

    2017年   岡山大学工学部  

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  • 内山勇三科学技術賞

    2016年  

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    受賞国:日本国

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  • The Journal of Biochemistry/OUP Poster Prize

    2009年  

     詳細を見る

    受賞国:日本国

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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年  

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    受賞国:日本国

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  • 生体機能関連化学シンポジウム講演賞

    2007年  

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    受賞国:日本国

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共同研究・競争的資金等の研究

  • 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年04月 - 2022年03月

    科研費 基盤研究(B) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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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年04月 - 2019年03月

    科研費 挑戦的研究(萌芽) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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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年04月 - 2017年03月

    科研費 挑戦的萌芽研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • 光増感剤修飾分子を用いたPCI の分子科学

    2014年04月 - 2016年03月

    科研費 新学術領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • PCDR技術による細胞機能の光制御

    2013年04月 - 2016年03月

    科研費 基盤研究(B) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • Sonochemical internalization法の確立

    2013年04月 - 2015年03月

    科研費 挑戦的萌芽研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • 光増感によるエンドソーム脱出の分子科学

    2012年04月 - 2014年03月

    科研費 新学術領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • RNAが引き起こす細胞内イベントの細胞周期依存性の解析

    2011年04月 - 2013年03月

    科研費 挑戦的萌芽研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • CLIP-RNAi法の開発と応用

    2009年04月 - 2012年03月

    科研費 若手研究(A) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • RNAi創薬を目指したRNAデリバリー技術の開発

    2008年04月 - 2010年03月

    科研費 特定領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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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

    2008年04月 - 2010年03月

    科研費 特定領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • 人工核酸によるRNAの検出と精製

    2007年04月 - 2009年03月

    科研費 特定領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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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

    2006年04月 - 2008年03月

    科研費 特定領域研究 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • RNAの新規細胞内導入法の開発と応用

    2006年01月 - 2008年12月

    NEDO  産業技術研究助成事業 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:48100000円

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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

    2005年04月 - 2007年03月

    科研費 若手研究(B) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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  • 異常tRNAを用いた翻訳系

    2002年04月 - 2003年10月

    科学技術振興調整費 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:34000000円 ( 直接経費:29130000円 )

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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

    2001年04月 - 2002年03月

    科研費 若手研究(B) 

    大槻 高史

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    担当区分:研究代表者  資金種別:競争的資金

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▼全件表示

 

担当授業科目

  • バイオ・創薬科学概論 (2021年度) 前期  - その他

  • バイオ・創薬科学概論 (2021年度) 前期  - 木1~2

  • ヘルスシステム統合科学専門英語 (2021年度) 後期  - その他

  • ヘルスシステム統合科学専門英語 (2021年度) 後期  - その他

  • ヘルスシステム統合科学特別研究 (2021年度) 通年  - その他

  • ヘルスシステム統合科学特別研究 (2021年度) 通年  - その他

  • 化学・生命系入門 (2021年度) 第1学期  - 金1~2

  • 化学生物学 (2021年度) 後期  - その他

  • 技術表現発表学 (2021年度) 後期  - その他

  • 技術表現発表学 (2021年度) 後期  - その他

  • 物理化学3 (2021年度) 第1学期  - 火5,火6,金1,金2

  • 物理化学4 (2021年度) 第1学期  - 火5,火6,金1,金2

  • 生化学3 (2021年度) 第3学期  - 月3,月4,水1,水2

  • 生化学4 (2021年度) 第3学期  - 月3,月4,水1,水2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2021年度) 3・4学期  - 金1,金2

  • 生物学基礎1 (2021年度) 第3学期  - 金1,金2

  • 生物学基礎1 (2021年度) 第3学期  - 金1,金2

  • 生物学基礎1 (2021年度) 第3学期  - 金1,金2

  • 生物学基礎2 (2021年度) 第4学期  - 金1,金2

  • 生物学基礎2 (2021年度) 第4学期  - 金1,金2

  • 生物学基礎2 (2021年度) 第4学期  - 金1,金2

  • 遺伝子工学 (2021年度) 第4学期  - 火5,火6

  • 遺伝子工学 (2021年度) 第4学期  - 火5,火6

  • RNA工学 (2021年度) 前期  - 火5~6

  • バイオ・創薬科学概論 (2020年度) 前期  - 木1,木2

  • ヘルスシステム統合科学インターンシップ (2020年度) 通年  - その他

  • ヘルスシステム統合科学専門英語 (2020年度) 後期  - その他

  • ヘルスシステム統合科学特別研究 (2020年度) 通年  - その他

  • 化学生物学 (2020年度) 後期  - その他

  • 技術表現発表学 (2020年度) 後期  - その他

  • 放射線安全利用工学1 (2020年度) 第1学期  - 金5,金6

  • 放射線安全利用工学2 (2020年度) 第2学期  - 金5,金6

  • 放射線安全利用工学及び実験 (2020年度) 1・2学期  - 金5,金6

  • 物理化学3 (2020年度) 第1学期  - 火5,火6,金1,金2

  • 物理化学4 (2020年度) 第1学期  - 火5,火6,金1,金2

  • 生化学3 (2020年度) 第3学期  - 月3,月4,水1,水2

  • 生化学4 (2020年度) 第3学期  - 月3,月4,水1,水2

  • 生物学基礎 (2020年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2020年度) 3・4学期  - 金1,金2

  • 生物学基礎 (2020年度) 3・4学期  - 金1,金2

  • 生物学基礎1 (2020年度) 第3学期  - 金1,金2

  • 生物学基礎1 (2020年度) 第3学期  - 金1,金2

  • 生物学基礎1 (2020年度) 第3学期  - 金1,金2

  • 生物学基礎2 (2020年度) 第4学期  - 金1,金2

  • 生物学基礎2 (2020年度) 第4学期  - 金1,金2

  • 生物学基礎2 (2020年度) 第4学期  - 金1,金2

  • 遺伝子工学 (2020年度) 第4学期  - 火5,火6

  • 遺伝子工学 (2020年度) 第4学期  - 火5,火6

▼全件表示