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Engineering of translation molecules for synthetic biology approach on the genetic code

Research Project

Project/Area Number 18H02416
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Sakamoto Kensaku  国立研究開発法人理化学研究所, 生命機能科学研究センター, チームリーダー (50240685)

Project Period (FY) 2018-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥15,340,000 (Direct Cost: ¥11,800,000、Indirect Cost: ¥3,540,000)
Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Keywords拡張遺伝暗号 / 非天然アミノ酸 / tRNA / アミノアシルtRNA合成酵素 / タンパク質合成 / 抗体 / 抗原親和性 / Fab / メチルグルタミン / 酵素変異体 / 合成生物学 / タンパク質工学 / 翻訳後修飾 / 遺伝暗号 / 非天然型アミノ酸 / 古細菌 / ピロリジン / アミノアシル化 / 立体構造
Outline of Final Research Achievements

The field of ‘genetic code expansion’ or ‘genetic code engineering’ encompasses two directions: Technology developments for in vivo incorporation of non-natural amino acids into proteins and applications of this technology in medical and industrial spheres. The present study successfully addressed both aspects of the field. A new approach for creating ‘identityless’ tRNAs, which are of fundamental importance for expanding the code, was thus established. Another achievement was a novel method to improve the antigen-binding of antibodies, which illuminated a new direction of protein engineering based on expanded codes.

Academic Significance and Societal Importance of the Research Achievements

遺伝暗号は全ての生き物にとっての“OS(オペレーティングシステム)”と言っても過言ではなく、拡張遺伝暗号とはその遺伝暗号がバージョンアップされたものである。合成生物学とは、生体分子、微生物等における有用な新規機能を将来に渉って持続的に開発することを可能にする学問であり、本課題では、非天然アミノ酸をタンパク質に導入するための遺伝暗号技術を一層発展させるとともに、社会的ニーズに答えるための応用研究を推進した。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (17 results)

All 2022 2021 2019 2018

All Journal Article (10 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 7 results,  Open Access: 4 results) Presentation (7 results) (of which Invited: 4 results)

  • [Journal Article] The two domain architecture of LAMP2A regulates its interaction with Hsc702022

    • Author(s)
      Yuta Ikami, Kazue Terasawa, Kensaku Sakamoto, Kazumasa Ohtake, Hiroyuki Harada Tetsuro Watabe, Shigeyuki Yokoyama, Miki Hara-Yokoyam
    • Journal Title

      Experimental Cell Research

      Volume: 411 Issue: 1 Pages: 112986-112986

    • DOI

      10.1016/j.yexcr.2021.112986

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] An integrated computational pipeline for designing high-affinity nanobodies with expanded genetic codes2021

    • Author(s)
      Aditya K Padhi, Ashutosh Kumar, Ken-ichi Haruna, Haruna Sato, Hiroko Tamura, Satoru Nagatoishi, Kouhei Tsumoto, Atushi Yamaguchi, Fumie Iraha, Mihoko Takahashi, Kensaku Sakamoto, Kam Y J Zhang
    • Journal Title

      Briefings in Bioinformatics

      Volume: 22 Issue: 6

    • DOI

      10.1093/bib/bbab338

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] 生細胞における遺伝暗号の改変と応用2021

    • Author(s)
      坂本健作
    • Journal Title

      生化学

      Volume: 93 Pages: 291-297

    • Related Report
      2021 Annual Research Report
  • [Journal Article] Direct homophilic interaction of LAMP2A with the two-domain architecture revealed by site-directed photo-crosslinks and steric hindrances in mammalian cells2021

    • Author(s)
      Terasawa Kazue、Kato Yuji、Ikami Yuta、Sakamoto Kensaku、Ohtake Kazumasa、Kusano Seisuke、Tomabechi Yuri、Kukimoto-Niino Mutsuko、Shirouzu Mikako、Guan Jun-Lin、Kobayashi Toshihide、Iwata Takanori、Watabe Tetsuro、Yokoyama Shigeyuki、Hara-Yokoyama Miki
    • Journal Title

      Autophagy

      Volume: 17 Issue: 12 Pages: 1-19

    • DOI

      10.1080/15548627.2021.1911017

    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] An expanded genetic code facilitates antibody chemical conjugation involving the lambda light chain2021

    • Author(s)
      Kato Akifumi、Ohtake Kazumasa、Tanaka Yoshitaka、Yokoyama Shigeyuki、Sakamoto Kensaku、Shiraishi Yasuhisa
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 546 Pages: 35-39

    • DOI

      10.1016/j.bbrc.2021.02.005

    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Incorporation of Halogenated Amino Acids into Antibody Fragments at Multiple Specific Sites Enhances Antigen Binding2021

    • Author(s)
      Akiko Hayashi Dr. Ken‐ichi Haruna Haruna Sato Dr. Kenichiro Ito Dr. Chisato Makino Dr. Takuhiro Ito Dr. Kensaku Sakamoto
    • Journal Title

      ChemBioChem

      Volume: 22 Issue: 1 Pages: 120-123

    • DOI

      10.1002/cbic.202000429

    • Related Report
      2021 Annual Research Report 2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis for genetic-code expansion with bulky lysine derivatives by an engineered pyrrolysyl-tRNA synthetase2019

    • Author(s)
      Tatsuo Yanagisawa, Mitsuo Kuratani, Eiko Seki, Nobumasa Hino, Kensaku Sakamoto, Shigeyuki Yokoyama
    • Journal Title

      Cell Chemical Biology

      Volume: 26 Pages: 936-949

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthetic tyrosine tRNA molecules with noncanonical secondary structures2019

    • Author(s)
      K. Sakamoto, A. Hayashi
    • Journal Title

      Int. J. Mol. Sci.

      Volume: 20 Issue: 1 Pages: 92-92

    • DOI

      10.3390/ijms20010092

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Cell-free protein synthesis using S30 extracts from Escherichia coli RFzero strains for efficient incorporation of non-natural amino acids into proteins2019

    • Author(s)
      J. Adachi, K. Katsura, E. Seki, C. Takemoto, M. Shirouzu, T. Terada, T. Mukai, K. Sakamoto, S. Yokoyama
    • Journal Title

      Int. J. Mol. Sci.

      Volume: 20 Issue: 3 Pages: 492-492

    • DOI

      10.3390/ijms20030492

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Pyrrolysyl-tRNA synthetase with a unique architecture enhances the availability of lysine derivatives in synthetic genetic codes2018

    • Author(s)
      A. Yamaguchi, F. Iraha, K. Ohtake, K. Sakamoto
    • Journal Title

      Molcules

      Volume: 23 Issue: 10 Pages: 2460-2460

    • DOI

      10.3390/molecules23102460

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 非アミノ酸を含む遺伝暗号の開発と大腸菌進化への影響2022

    • Author(s)
      大竹和正, 芝井厚, 古澤力, 坂本健作
    • Organizer
      第16回無細胞生命化学研究会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 遺伝暗号改変が拓くタンパク質工学2021

    • Author(s)
      坂本健作
    • Organizer
      日本薬学会第141年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 10.非天然型アミノ酸含有抗体発現生産技術と生体直交反応性キレーター修飾の組み合わせによる高均一な64Cu標識トラスツズマブFabの作製2019

    • Author(s)
      仁欽、藁科翔太、毛利浩太、高橋麻衣子、金山洋介、高橋美穂子、林明子、林中恵美、和田康弘、渡辺恭良、坂本健作、向井英史
    • Organizer
      日本分子イメージング学会第14回学術総会・学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Xeno-protein engineeringの抗体開発への応用2019

    • Author(s)
      坂本健作、林明子、山口純、柳沢達男、田村浩子、長門石曉、津本浩平、藏地理代、金山洋介、渡辺恭良
    • Organizer
      第14回無細胞生命科学研究会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 人工遺伝暗号技術の抗体医薬への応用2019

    • Author(s)
      坂本健作
    • Organizer
      理研シンポジウム「理研エンジニアリングネットワーク/精密武装抗体の合成と機能評価」
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 改変型抗体の作製のための技術基盤2019

    • Author(s)
      坂本健作
    • Organizer
      兵庫県立こども病院-理研BDR第4回ジョイントシンポジウム
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] ゼノバイオロジーを目指すタンパク質工学2019

    • Author(s)
      坂本健作
    • Organizer
      日本薬学会第140年会シンポジウム
    • Related Report
      2019 Annual Research Report
    • Invited

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Published: 2018-04-23   Modified: 2023-01-30  

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