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Photo-triggered translation using caged aminoacyl-tRNA in early embryos

Research Project

Project/Area Number 17K19202
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Research Field Biomolecular chemistry and related fields
Research InstitutionOkayama University

Principal Investigator

Ohtsuki Takashi  岡山大学, ヘルスシステム統合科学研究科, 教授 (80321735)

Co-Investigator(Kenkyū-buntansha) 若井 拓哉  岡山大学, 環境生命科学研究科, 准教授 (60557768)
Project Period (FY) 2017-06-30 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
KeywordsケージドアミノアシルtRNA / 翻訳 / 光制御 / 発生
Outline of Final Research Achievements

Light is one of the most easily manipulated external factors for spatiotemporally controlling biological events, such as protein biosynthesis. We synthesized a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-caged aminoacyl-tRNAs (aa-tRNAs) for photocontrolling protein biosynthesis. Upon irradiation at 400-430 nm, DEACM-aa-tRNA was rapidly converted into active aa-tRNA. Translation was photoinduced when DEACM-aa-tRNAs carrying a CUA anticodon were used together with mRNAs harboring a UAG amber codon. Protein synthesis was phototriggered not only in an in vitro translation system, but also in mammalian cells. We tried to apply this strategy to photo-trigger synthesis of a fluorescent protein and PGC-1alpha in a mouse embryonic cell.

Academic Significance and Societal Importance of the Research Achievements

本研究で行ったことは、特定のタンパク質の合成(発現)を時空間的に制御する方法の開発である。マウス胚において、効率よく特異的な発現をさせるためには、まだ改善を要することが分かったが、胚を含めて動物個体や三次元的な細胞集合体に適用可能になれば、“時間的”翻訳制御と“空間的”翻訳制御の双方の側面を生かして、様々なタンパク質の役割の解明が可能になる。また、光による時空間的タンパク質発現制御を通じて、初期発生事象の研究および人為的コントロールも可能になると期待される。

Report

(3 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • Research Products

    (11 results)

All 2019 2018 2017

All Journal Article (4 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 4 results) Presentation (6 results) (of which Invited: 1 results) Book (1 results)

  • [Journal Article] Constitutive IP3R1-mediated Ca2+ release reduces Ca2+ store content and stimulates mitochondrial metabolism in mouse GV oocytes2019

    • Author(s)
      Wakai T, Fissore RA
    • Journal Title

      Journal of Cell Science

      Volume: 132 Issue: 3 Pages: 1-1

    • DOI

      10.1242/jcs.225441

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Red and Near-Infrared Light-Directed Cytosolic Delivery of Two Different RNAs Using Photosensitive RNA Carriers2018

    • Author(s)
      Shiraga Kaori、Soe Tet Htut、Matsumoto Sho、Watanabe Kazunori、Ohtsuki Takashi
    • Journal Title

      Bioconjugate Chemistry

      Volume: 29 Issue: 9 Pages: 3174-3179

    • DOI

      10.1021/acs.bioconjchem.8b00487

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enhanced intracellular peptide delivery by multivalent cell-penetrating peptide with bioreducible linkage.2017

    • Author(s)
      Kim, H., Kitamatsu, M., Ohtsuki, T.
    • Journal Title

      Bioorganic & Medicinal Chemistry Letters

      Volume: 28 Issue: 3 Pages: 378-381

    • DOI

      10.1016/j.bmcl.2017.12.035

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles.2017

    • Author(s)
      Bui TMT, Nguyen KX, Karata A, Ferre P, Tran MT, Wakai T, Funahashi H.
    • Journal Title

      Reproduction, Fertility and Development.

      Volume: 29 Issue: 10 Pages: 1902-1909

    • DOI

      10.1071/rd16321

    • NAID

      120006348281

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Presentation] 光依存的なRNA導入法による2種類のRNAの時間差導入2018

    • Author(s)
      大槻高史, 白神 かおり, 渡邉 和則
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 光に応答し細胞質内に侵入する機能性光増感剤ペプチドの設計2018

    • Author(s)
      三好 祐一, 山本 怜見, 門野 真保, 北松 瑞生, 渡邉 和則, 大槻 高史
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] マウス卵母細胞におけるTrim-away法を用いたDrp1タンパク質の分解2018

    • Author(s)
      月向はるな、野村瑠莉、舟橋 弘晃、若井 拓哉
    • Organizer
      第18回日本ミトコンドリア学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 着床前マウス胚におけるミトコンドリア分裂因子Drp1の役割2018

    • Author(s)
      野村瑠莉、月向はるな、舟橋弘晃、若井拓哉
    • Organizer
      第18回日本ミトコンドリア学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ケージドアミノアシルtRNAを用いたタンパク質生合成の光誘導2017

    • Author(s)
      大槻高史、神崎重人、西村紗恵、国広芳朗、渡邉和則
    • Organizer
      第11回バイオ関連化学シンポジウム
    • Related Report
      2017 Research-status Report
  • [Presentation] 卵母細胞における小胞体Ca2+ の制御とその役割2017

    • Author(s)
      若井拓哉
    • Organizer
      第35回日本受精着床学会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Book] Encyclopedia of Reproduction (Second Edition)2018

    • Author(s)
      Wakai T., Mehregan A., Fissore R.A.(部分執筆)
    • Total Pages
      3868
    • Publisher
      Elsevier
    • ISBN
      9780128118993
    • Related Report
      2018 Annual Research Report

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Published: 2017-07-21   Modified: 2020-03-30  

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