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Clarification of Cas9"s target DNA recognition mechanisim.

Research Project

Project/Area Number 17K15100
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionThe University of Tokyo

Principal Investigator

Komori Tomotaka  東京大学, 大学院理学系研究科(理学部), 客員共同研究員 (50542289)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2017: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Keywordsゲノム編集タンパク質 / 1分子計測 / FRET / ゲノム編集 / 蛍光計測 / ダイナミクス / 核酸結合タンパク質 / 1分子計測 / 構造ゆらぎ / 生物物理 / バイオテクノロジー / 遺伝子
Outline of Final Research Achievements

Cas9 bound with single-guide RNA (sgRNA) cleaves its target DNA sequence depending on the sgRNA's sequence. Because of this Cas9's property, Cas9 is widely utilized for the genome editing tool. In this study, we revealed using single-molecule FRET technique that dynamic fluctuation of HNH domain (Cas9"s nuclease domain) is important for DNA cleavage activity of Cas9.

Academic Significance and Societal Importance of the Research Achievements

Cas9はsgRNAの配列を変更する事で任意のターゲットDNA配列の切断が可能である事から近年、ゲノム編集のためのツールとして広く利用されている。これまでにCas9がDNA配列を切断する際の詳細な分子機構は解明されていなかった。本研究では1分子FRET法を用いる事でCas9がターゲットDNA配列を切断する際のCas9内部の動的な構造変化を明らかにした。本研究で得られた分子機構を基盤とすることで、本成果は将来のより効率的なCas9の改良へと繋がる事が期待出来る。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (8 results)

All 2018 2017

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (7 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Real-time observation of flexible movements in CRISPR-Cas92018

    • Author(s)
      S. Osuka, K. Isomura, S. Kajimoto, T. Komori, H. Nishimasu, T. Shima, O. Nureki, S. Uemura
    • Journal Title

      EMBO Journal

      Volume: 印刷中 Issue: 10

    • DOI

      10.15252/embj.201796941

    • Related Report
      2018 Research-status Report 2017 Research-status Report
    • Peer Reviewed
  • [Presentation] N-terminal residues of Drosophila Argonaute2 possess the ability to form amyloid fibrils2018

    • Author(s)
      成田 晴香, 桑原 誠, 小森 智貴, 村上 僚, 島 知弘, 塩見 美喜子, 上村 想太郎
    • Organizer
      日本蛋白質科学会第18回年会
    • Related Report
      2018 Research-status Report
  • [Presentation] N-terminal residues of Drosophila Argonaute2 posesses the abilty to form amyloid fibrils.2018

    • Author(s)
      Haruka Narita, Makoto F Kuwabara, Tomotaka Komori, Ryo Murakami, Tomohiro Shima, Mikiko C Siomi, Sotaro Uemura
    • Organizer
      Asian Pacific Prion Society 2018
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] N-terminal residues of Drosophila Argonaute2 possess the ability to form amyloid fibrils2018

    • Author(s)
      成田 晴香, 桑原 誠, 小森 智貴, 村上 僚, 島 知弘, 塩見 美喜子, 上村 想太郎
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 電位駆動型モータープレスチン以外の SLC26 イオン輸送体にも電位感受能は存在する2018

    • Author(s)
      桑原誠、和佐野浩一郎、高橋里枝、Justin Bodner、小森智貴、上村想太郎、Jing Zheng、島知弘、本 間和明
    • Organizer
      日本蛋白質科学会第18回年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Reconstitution system of Siwi- and Vasa- coupled piRNA biogenesis2018

    • Author(s)
      Tomotaka Komori, Ryo Murakami, Tomohiro Shima, Mikiko C. Siomi, Sotaro Uemura
    • Organizer
      日本蛋白質科学会第18回年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Reconstitution system of Siwi- and Vasa-coupled piRNA biogenesis.2017

    • Author(s)
      Tomotaka Komori, Ryo Murakami, Tomohiro Shima, Mikiko C Siomi, Sotaro
    • Organizer
      日本生物物理学会第55回年会
    • Related Report
      2017 Research-status Report
  • [Presentation] Single molecule pathway monitoring of the Argonaute RISC formation process.2017

    • Author(s)
      T. Komori, R. Murakami, M. Moriya,K.M. Nishida, M.C. Siomi, S. Uemura
    • Organizer
      The 43rd Naito Conference
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research

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Published: 2017-04-28   Modified: 2021-02-19  

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