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Rational construction of eukaryotic riboswitches requiring less energy for their conformational changes

Research Project

Project/Area Number 16K05846
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Bio-related chemistry
Research InstitutionEhime University

Principal Investigator

Ogawa Atsushi  愛媛大学, プロテオサイエンスセンター, 准教授 (30442940)

Project Period (FY) 2016-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsリボスイッチ / 発現制御 / 合成生物学 / 翻訳促進 / IRES / CITE
Outline of Final Research Achievements

We rationally constructed three types of energy-saving, eukaryotic riboswitches (ligand-responsive regulatory RNAs) that function in eukaryotic translation systems. The first one upregulates translation mediated by a 3' cap-independent translation element (3' CITE) in response to its specific ligand. The second one ligand-dependently downregulates translation mediated by an internal ribosome entry site (IRES). The last one ligand-responsively downregulates canonical translation. Because all of them require no major hybridization switches for gene regulation, they require less energy for their conformational changes, thus showing higher switching efficiencies. In addition, their ligand-specificity can easily be changed just by replacing a part of them (specifically, the aptamer domain) to obtain a riboswitch responsive to a user-defined ligand.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した省エネ型リボスイッチは、分子応答制御時に「鎖置換」を伴わないため、エネルギーロスが少なく、高い制御効率を発揮する。また、分子特異性の変更も容易であることから、任意の分子の濃度に応じて任意のタンパク質の生産量を広範囲にコントロールすることが可能となる。このような人工の分子応答性遺伝子発現制御システムは、合成生物学を含む幅広い学問領域において、基盤技術としての利用が期待される。

Report

(5 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (28 results)

All 2019 2018 2017 2016 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (19 results) (of which Int'l Joint Research: 8 results,  Invited: 1 results) Book (1 results) Remarks (1 results)

  • [Journal Article] Detection of Gold Nanoparticles Aggregation Using Light Scattering for Molecular Sensing2019

    • Author(s)
      Yuki Yano, Masamichi Nisougi, Yuki Yano-Ozawa, Tsuyoshi Ohguni, Atsushi Ogawa, Mizuo Maeda, Tsuyoshi Asahi, Tamotsu Zako
    • Journal Title

      Analytical Sciences

      Volume: 35 Issue: 6 Pages: 685-690

    • DOI

      10.2116/analsci.18P571

    • NAID

      130007661696

    • ISSN
      0910-6340, 1348-2246
    • Year and Date
      2019-06-10
    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] In vitro selection of a 3′ terminal short protector that stabilizes transcripts to improve the translation efficiency in a wheat germ extract2019

    • Author(s)
      Atsushi Ogawa, Akane Kutsuna, Masashi Takamatsu, Tatsuya Okuzono
    • Journal Title

      Bioorganic & Medicinal Chemistry Letters

      Volume: 29 Issue: 16 Pages: 2141-2144

    • DOI

      10.1016/j.bmcl.2019.06.058

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mutation of the start codon to enhance Cripavirus internal ribosome entry site-mediated translation in a wheat germ extract2019

    • Author(s)
      Atsushi Ogawa and Masashi Takamatsu
    • Journal Title

      Bioorganic & Medicinal Chemistry Letters

      Volume: 29 Issue: 22 Pages: 126729-126729

    • DOI

      10.1016/j.bmcl.2019.126729

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Modification of carbon metabolism in Synechococcus elongatus PCC 7942 by cyanophage-derived sigma factors for bioproduction improvement2019

    • Author(s)
      Naruhiko Sawa, Tsuneyuki Tatsuke, Atsushi Ogawa, Yasutaka Hirokawa,Takashi Osanai, Taizo Hanai
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 127 Issue: 2 Pages: 256-264

    • DOI

      10.1016/j.jbiosc.2018.07.019

    • NAID

      40021813937

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Canonical translation-modulating OFF-riboswitches with a single aptamer binding to a small molecule that function in a higher eukaryotic cell-free expression system2018

    • Author(s)
      Atsushi Ogawa, Yuta Murashige, Hajime Takahashi
    • Journal Title

      Bioorganic and Medicinal Chemistry Letters

      Volume: 28 Issue: 14 Pages: 2353-2357

    • DOI

      10.1016/j.bmcl.2018.06.041

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Artificial OFF-Riboswitches That Downregulate Internal Ribosome Entry without Hybridization Switches in a Eukaryotic Cell-Free Translation System2017

    • Author(s)
      Atsushi Ogawa, Hiroki Masuoka, Tsubasa Ota
    • Journal Title

      ACS Synthetic Biology

      Volume: 6 Issue: 9 Pages: 1656-1662

    • DOI

      10.1021/acssynbio.7b00124

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Ligand-responsive upregulation of 3' CITE-mediated translation in a wheat germ cell-free expression system2017

    • Author(s)
      Atsushi Ogawa, Yuta Murashige, Junichiro Tabuchi, Taiki Omatsu
    • Journal Title

      Molecular BioSystems

      Volume: 13 Issue: 2 Pages: 314-319

    • DOI

      10.1039/c6mb00748a

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Gene expression in a super-giant liposome encapsulating a wheat germ extract2019

    • Author(s)
      Hajime Takahashi, Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ナノDNA応答性真核系人工リボスイッチの構築2019

    • Author(s)
      伊藤 優, 小川 敦司
    • Organizer
      「細胞を創る」研究会12.0
    • Related Report
      2019 Annual Research Report
  • [Presentation] リボスイッチ媒介化学コミュニケーション型の真核系超巨大人工細胞2019

    • Author(s)
      髙橋 萌, 小川 敦司
    • Organizer
      「細胞を創る」研究会12.0
    • Related Report
      2019 Annual Research Report
  • [Presentation] EVALUATION OF THEOPHYLLINE-DEPENDENT SYNTHETIC RIBOSWITCHES FOR STRICT CONTROL OF PLASTID GENE EXPRESSION2018

    • Author(s)
      Rika Yamane, Atsushi Ogawa, Yuzuru Tozawa, Takashi Shiina, Yoichi Nakahira
    • Organizer
      INTERNATIONAL PLANT MOLECULAR BIOLOGY 2018
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] In vitro selection of a 3’ protector that stabilizes mRNA to increase the translation efficiency in a wheat germ cell-free expression system2018

    • Author(s)
      Akane Kutsuna, Tatsuya Okuzono, Masashi Takamatsu, Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2018
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Artificial OFF-riboswitches that downregulate internal ribosome entry without hybridization switches in a wheat germ extract2018

    • Author(s)
      Hiroki Masuoka, Tsubasa Ota, Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2018
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] コムギ無細胞系におけるmRNA安定化および翻訳効率上昇を指向した3’末端保護配列のin vitro selection2018

    • Author(s)
      忽那 茜、奥園 達也、高松 将史、小川 敦司
    • Organizer
      第13回無細胞生命科学研究会
    • Related Report
      2018 Research-status Report
  • [Presentation] 省エネ型・IRES基盤真核系人工OFFリボスイッチの開発2018

    • Author(s)
      升岡 宏紀、太田 翼、髙橋 萌、小川 敦司
    • Organizer
      第13回無細胞生命科学研究会
    • Related Report
      2018 Research-status Report
  • [Presentation] 鎖置換反応を必要としないIRES基盤真核系人工OFFリボスイッチの開発2018

    • Author(s)
      升岡 宏紀、太田 翼、髙橋 萌、小川 敦司
    • Organizer
      2018年日本化学会中国四国支部大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 小麦無細胞系におけるRNA3’末端保護配列のin vitro selection2018

    • Author(s)
      忽那 茜、奥園 達也、高松 将史、小川 敦司
    • Organizer
      2018年日本化学会中国四国支部大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 人工リボスイッチを基盤とした葉緑体遺伝子発現誘導系の改良2018

    • Author(s)
      山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Development of a strictly controlled inducible gene expression system for plastids by using synthetic riboswitches2018

    • Author(s)
      Rika Yamane, Atsushi Ogawa, Yuzuru Tozawa, Takashi Shiina, Yoichi Nakahira
    • Organizer
      Gordon Research Conference (Chloroplast Biotechnology 2019)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Rational design of artificial riboswitches that function in a eukaryotic expression system2017

    • Author(s)
      Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Ligand-responsive upregulation of 3' CITE-mediated translation in wheat germ extract2017

    • Author(s)
      Yuta Murashige, Junichiro Tabuchi, Taiki Omatsu, Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 人工リボスイッチを活用した葉緑体遺伝子発現誘導系の評価2017

    • Author(s)
      山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 人工リボスイッチを活用した葉緑体遺伝子発現制御系の評価2017

    • Author(s)
      山根 里佳、小川 敦司、戸澤 譲、椎名 隆、中平 洋一
    • Organizer
      第35回日本植物細胞分子生物学会
    • Related Report
      2017 Research-status Report
  • [Presentation] 鎖置換を利用しない真核系人工ONリボスイッチの合理設計2016

    • Author(s)
      村重 裕太、田渕 潤一郎、大松 大希、小川 敦司
    • Organizer
      「細胞を創る」研究会9.0
    • Place of Presentation
      早稲田大学(東京都・新宿区)
    • Year and Date
      2016-11-21
    • Related Report
      2016 Research-status Report
  • [Presentation] Facile design of artificial riboswitches that ligand-dose dependently upregulate translation without hybridization switches in a wheat germ extract2016

    • Author(s)
      Yuta Murashige, Junichiro Tabuchi, Taiki Omatsu, Atsushi Ogawa
    • Organizer
      Protein Island Matsuyama International Symposium 2016,
    • Place of Presentation
      愛媛大学(愛媛県・松山市)
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] 省エネ型真核リボスイッチの設計2016

    • Author(s)
      小川 敦司
    • Organizer
      新世代の生物有機化学研究会2016
    • Place of Presentation
      愛媛大学(愛媛県・松山市)
    • Related Report
      2016 Research-status Report
  • [Book] Applied RNA Bioscience2018

    • Author(s)
      Seiji Masuda, Shingo Izawa, Atsushi Ogawa他
    • Total Pages
      286
    • Publisher
      Springer
    • ISBN
      9789811083716
    • Related Report
      2017 Research-status Report
  • [Remarks] 愛媛大学 小川研究室

    • URL

      http://www.pros.ehime-u.ac.jp/ogawa/

    • Related Report
      2019 Annual Research Report 2018 Research-status Report 2017 Research-status Report 2016 Research-status Report

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Published: 2016-04-21   Modified: 2021-02-19  

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