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Directed Evolution of Nucleic Acid Enzymes Combining Experiment and Computation

Research Project

Project/Area Number 19H02855
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 37030:Chemical biology-related
Research InstitutionOkinawa Institute of Science and Technology Graduate University

Principal Investigator

Yokobayashi Yohei  沖縄科学技術大学院大学, 核酸化学・工学ユニット, 教授 (70769752)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2021: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2020: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2019: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Keywords進化工学 / 核酸化学 / RNA / リボザイム / 核酸酵素 / 合成生物学
Outline of Research at the Start

遺伝子情報を担うDNAやRNAといった核酸は、タンパク質酵素のように化学反応を触媒する機能も持ち、物質生産や遺伝子治療などに応用が期待されている。本研究の主な目的は、これらの核酸酵素を実験室内で人工的に進化させ、より活性の高い配列を発見することである。さらに、この人工進化の過程に、機械学習などコンピューターによる計算を取り入れ、従来の実験室進化を凌駕する手法を開発することを目指す。

Outline of Final Research Achievements

Catalytic nucleic acids, especially those based on RNAs (ribozymes), are intriguing molecules for their implications in the origin of life and applications in gene therapy. The main objective of this project is to measure and predict the sequence-function relationships of these ribozymes. In this project, we measured activities of a large set of ribozyme sequences that have RNA ligase activity. The data were then analyzed by various methods including machine learning. As a result, we obtained various insights into the relationships between ribozyme sequence and its function.

Academic Significance and Societal Importance of the Research Achievements

触媒機能をもつRNA、リボザイムは、生命の起源に深く関わると考えられている。また、遺伝子治療などへの応用も有望な機能性核酸である。本研究は大量のリボザイム塩基配列とその機能を実験的に測定し、得られた大量のデータを機械学習などで解析することにより、これまでにない基礎的な知見が得られた。本研究により開発されたリボザイム測定技術やデータ解析法は、他の核酸酵素にも応用可能であり、様々な発展が期待できる。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (13 results)

All 2022 2021 2020 2019

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 6 results) Presentation (6 results) (of which Int'l Joint Research: 4 results,  Invited: 4 results)

  • [Journal Article] High-throughput screening of cell-free riboswitches by fluorescence-activated droplet sorting2022

    • Author(s)
      Tabuchi Takeshi、Yokobayashi Yohei
    • Journal Title

      Nucleic Acids Research

      Volume: 50 Issue: 6 Pages: 3535-3550

    • DOI

      10.1093/nar/gkac152

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Directed evolution of orthogonal RNA-RBP pairs through library-vs-library in vitro selection2022

    • Author(s)
      Keisuke Fukunaga and Yohei Yokobayashi
    • Journal Title

      Nucleic Acids Research

      Volume: 50 Issue: 2 Pages: 601-616

    • DOI

      10.1093/nar/gkab527

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Circularly-Permuted Pistol Ribozyme: A Synthetic Ribozyme Scaffold for Mammalian Riboswitches2021

    • Author(s)
      Mustafina Kamila、Nomura Yoko、Rotrattanadumrong Rachapun、Yokobayashi Yohei
    • Journal Title

      ACS Synthetic Biology

      Volume: 10 Issue: 8 Pages: 2040-2048

    • DOI

      10.1021/acssynbio.1c00213

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] High-Throughput Analysis and Engineering of Ribozymes and Deoxyribozymes by Sequencing2020

    • Author(s)
      Yokobayashi Yohei
    • Journal Title

      Accounts of Chemical Research

      Volume: 53 Issue: 12 Pages: 2903-2912

    • DOI

      10.1021/acs.accounts.0c00546

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Design of Mammalian ON-Riboswitches Based on Tandemly Fused Aptamer and Ribozyme2020

    • Author(s)
      Kamila Mustafina, Keisuke Fukunaga, and Yohei Yokobayashi
    • Journal Title

      ACS Synthetic Biology

      Volume: 9 Issue: 1 Pages: 19-25

    • DOI

      10.1021/acssynbio.9b00371

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Self-powered RNA nanomachine driven by metastable structure2019

    • Author(s)
      Kobori Shungo、Nomura Yoko、Yokobayashi Yohei
    • Journal Title

      Nucleic Acids Research

      Volume: 47 Issue: 11 Pages: 6007-6014

    • DOI

      10.1093/nar/gkz364

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Systematic minimization of RNA ligase ribozyme through large-scale design-synthesis-sequence cycles2019

    • Author(s)
      Nomura Yoko、Yokobayashi Yohei
    • Journal Title

      Nucleic Acids Research

      Volume: 47 Issue: 17 Pages: 8950-8960

    • DOI

      10.1093/nar/gkz729

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] High-throughput analysis of ribozyme sequence-function relationship2022

    • Author(s)
      Yohei Yokobayashi
    • Organizer
      RIKEN BDR Symposium 2022
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] セルフリーシステムにおける人工リボスイッチの構築2020

    • Author(s)
      横林 洋平
    • Organizer
      生物工学Webシンポジウム2020
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Designing RNA Devices in Test Tubes and Cells2019

    • Author(s)
      Yokobayashi Y.
    • Organizer
      46th International Symposium on Nucleic Acids Chemistry
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 人工細胞でも使える(?)RNAデバイスの作成2019

    • Author(s)
      横林 洋平
    • Organizer
      「細胞を創る」研究会12.0
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Systematic Minimization of RNA Ligase Ribozyme Through Large-Scale Design-Synthesis-Sequence Cycles2019

    • Author(s)
      Nomura Y, Yokobayashi Y.
    • Organizer
      46th International Symposium on Nucleic Acids Chemistry
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Design of Mammalian ON-riboswitches Based on Tandemly Fused Aptamer and Ribozyme2019

    • Author(s)
      Mustafina k, Fukunaga K, Yokobayashi Y.
    • Organizer
      46th International Symposium on Nucleic Acids Chemistry
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research

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Published: 2019-04-18   Modified: 2023-01-30  

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