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Development of genetically-encoded multi-color biosensors for intracellular signaling

Research Project

Project/Area Number 16K01922
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biomolecular chemistry
Research InstitutionTokyo Institute of Technology (2017-2019)
Waseda University (2016)

Principal Investigator

Kitaguchi Tetsuya  東京工業大学, 科学技術創成研究院, 准教授 (60432374)

Co-Investigator(Kenkyū-buntansha) 坪井 貴司  東京大学, 大学院総合文化研究科, 教授 (80415231)
上田 宏  東京工業大学, 科学技術創成研究院, 教授 (60232758)
Project Period (FY) 2016-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords蛍光タンパク質 / グルコース / バイオセンサー / バイオイメージング / GFP
Outline of Final Research Achievements

I have succeeded in developing genetically-encoded intensiometric sensors based on fluorescent proteins. These sensors enable us to visualize the intracellular dynamics of signaling molecules such as cGMP and cAMP, and metabolic molecules such as ATP and glucose by real time live-cell imaging. Furthermore, since these sensors have multiple colors such as green, red, and blue, several molecules can be simultaneously visualized by multi-color imaging. Therefore, they are able to clarify interactions and hierarchical relationships between intracellular molecules.

Academic Significance and Societal Importance of the Research Achievements

これまで単色で可視化できる遺伝子コード型センサーの種類は少なく、マルチカラーイメージングによる複数分子の同時解析は困難であった。本研究課題で開発した、cGMP、cAMP、ATP、グルコースのセンサーはマルチカラーイメージングに適用できるため、学術的意義は大きいと考えられる。また、センサー開発過程で蓄積したノウハウにより、以前よりシステマティックにセンサー開発ができるようになった。センサーは社会生活のさまざまな場面で利用できるため、センサーが自由自在に作出できることは社会的意義も大きい。

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

    (30 results)

All 2019 2018 2017 2016 Other

All Int'l Joint Research (4 results) Journal Article (7 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 5 results,  Open Access: 5 results,  Acknowledgement Compliant: 1 results) Presentation (17 results) Patent(Industrial Property Rights) (2 results) (of which Overseas: 2 results)

  • [Int'l Joint Research] National University of Singapore/Temasek Life Sciences Laboratory(シンガポール)

    • Related Report
      2018 Research-status Report
  • [Int'l Joint Research] Harvard University(米国)

    • Related Report
      2018 Research-status Report
  • [Int'l Joint Research] ASTAR/National University of Singapore(シンガポール)

    • Related Report
      2017 Research-status Report
  • [Int'l Joint Research] Harvard University(米国)

    • Related Report
      2017 Research-status Report
  • [Journal Article] Green Fluorescent Protein-Based Glucose Indicators Report Glucose Dynamics in Living Cells2019

    • Author(s)
      Mita Marie、Ito Motoki、Harada Kazuki、Sugawara Izumi、Ueda Hiroshi、Tsuboi Takashi、Kitaguchi Tetsuya
    • Journal Title

      Analytical Chemistry

      Volume: 91 Issue: 7 Pages: 4821-4830

    • DOI

      10.1021/acs.analchem.9b00447

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] RGB-Color Intensiometric Indicators to Visualize Spatiotemporal Dynamics of ATP in Single Cells2018

    • Author(s)
      Arai Satoshi、Kriszt R?kus、Harada Kazuki、Looi Liang-Sheng、Matsuda Shogo、Wongso Devina、Suo Satoshi、Ishiura Shoichi、Tseng Yu-Hua、Raghunath Michael、Ito Toshiro、Tsuboi Takashi、Kitaguchi Tetsuya
    • Journal Title

      Angewandte Chemie International Edition

      Volume: 57 Issue: 34 Pages: 10873-10878

    • DOI

      10.1002/anie.201804304

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Flashbody: A Next Generation Fluobody with Fluorescence Intensity Enhanced by Antigen Binding2017

    • Author(s)
      Wongso Devina、Dong Jinhua、Ueda Hiroshi、Kitaguchi Tetsuya
    • Journal Title

      Analytical Chemistry

      Volume: 89 Issue: 12 Pages: 6719-6725

    • DOI

      10.1021/acs.analchem.7b00959

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging2017

    • Author(s)
      Harada Kazuki、Ito Motoki、Wang Xiaowen、Tanaka Mika、Wongso Devina、Konno Ayumu、Hirai Hirokazu、Hirase Hajime、Tsuboi Takashi、Kitaguchi Tetsuya
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 7351-7351

    • DOI

      10.1038/s41598-017-07820-6

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] 蛍光蛋白質センサーをテーラーメード化2017

    • Author(s)
      北口哲也
    • Journal Title

      日経バイオテク

      Volume: 7月24日号 Pages: 42-45

    • Related Report
      2017 Research-status Report
  • [Journal Article] Generation of a cGMP indicator with an expanded dynamic range by optimization of amino acid linkers between a fluorescent protein and PDE5α.2017

    • Author(s)
      Matsuda S, Harada K, Ito M, Takizawa M, Wongso D, Tsuboi T, Kitaguchi T
    • Journal Title

      ACS Sensors

      Volume: 2 Issue: 1 Pages: 46-57

    • DOI

      10.1021/acssensors.6b00582

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] 蛍光・発光プローブ2016

    • Author(s)
      北口哲也
    • Journal Title

      光アライアンス

      Volume: 27 Pages: 15-19

    • Related Report
      2016 Research-status Report
  • [Presentation] 生細胞内のグルコース動態を検出する遺伝子コード型緑色蛍光センサーの創出2019

    • Author(s)
      三田真理恵、伊藤幹、原田一貴、菅原和、上田宏、坪井貴司、北口哲也
    • Organizer
      2019年度日本生化学会関東支部例会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 緑色蛍光グルコースセンサーが検出する人工甘味料による膵β細胞のグルコース恒常性攪乱2019

    • Author(s)
      三田真理恵、伊藤幹、原田一貴、菅原和、永田怜、芦田翼、上田宏、坪井貴司、北口哲也
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 蛍光タンパク質を用いた緑色蛍光輝度変化型乳酸センサーの開発2019

    • Author(s)
      千原貴美、三田真理恵、菅原和、相木彩希、早坂優希、原田一貴、上田宏、北口哲也、坪井貴司
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 生細胞イメージングを目指した遺伝子コード型ジベレリンプローブの開発2019

    • Author(s)
      相原寛史、Yanyan Hou、上田宏、伊藤寿朗、北口哲
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Single fluorescent protein-based sensors visualize dynamics of glucose by live cell imaging.2018

    • Author(s)
      Marie Mita, Motoki Ito, Kazuki Harada, Izumi Sugawara, Hiroshi Ueda, Takashi Tsuboi and Tetsuya Kitaguchi.
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 赤色蛍光タンパク質を用いたcGMP蛍光センサーの生細胞イメージングへの応用2018

    • Author(s)
      石田りか、三田真理恵、原田一貴、上田宏、北口哲也、坪井貴司
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 赤色蛍光グルコースセンサーによる細胞内グルコース動態の可視化2018

    • Author(s)
      菅原和、伊藤幹、三田真理恵、上田宏、坪井貴司、北口哲也
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 緑色蛍光タンパク質を用いたピルビン酸センサーの開発2018

    • Author(s)
      早坂優希、上田宏、北口哲也、坪井貴司
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Turn-on型の高輝度変化率を有する緑色cAMPセンサーの開発2018

    • Author(s)
      相木彩希、上田宏、北口哲也、坪井貴司
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 抗体を分子認識ドメインとする遺伝子コード型蛍光タンパク質プローブの開発2018

    • Author(s)
      相原寛史、Devina Wongso、董金華、上田宏、北口哲也
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] A red fluorescent protein-based cAMP indicator for dual color imaging, optogenetic manipulations and in vivo imaging.2017

    • Author(s)
      Kazuki Harada, Motoki Ito, Xiaowen Wang, Mika Tanaka, Devina Wongso, Hajime Hirase, Takashi Tsuboi and Tetsuya Kitaguchi.
    • Organizer
      第94回日本生理学会大会
    • Place of Presentation
      浜松
    • Year and Date
      2017-03-28
    • Related Report
      2016 Research-status Report
  • [Presentation] Flashbody: genetically-encoded antibody-based fluorescent probe with antigen dependent fluorescence intensity.2017

    • Author(s)
      Devina Wongso, Jinhua Dong, Hiroshi Ueda and Tetsuya Kitaguchi.
    • Organizer
      第17回蛋白質科学会
    • Related Report
      2017 Research-status Report
  • [Presentation] Hideki Itoh, Satoshi Arai, Thankiah Sudhaharan, Tetsuya Kitaguchi and E. Birgitte Lane. Single-cell quantitative analysis of ATP concentration by fluorescence lifetime imaging microscopy.2017

    • Author(s)
      Hideki Itoh, Satoshi Arai, Thankiah Sudhaharan, Tetsuya Kitaguchi and E. Birgitte Lane.
    • Organizer
      第55回日本生物物理学会
    • Related Report
      2017 Research-status Report
  • [Presentation] 蛍光タンパク質を用いた単色型グルコース可視化センサーの開発2017

    • Author(s)
      三田真理恵、伊藤幹、原田一貴、上田宏、坪井貴司、北口哲也
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 赤色蛍光タンパク質を用いたcGMP蛍光センサーの開発2017

    • Author(s)
      石田りか、三田真理恵、原田一貴、上田宏、北口哲也、坪井貴司
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 赤色蛍光タンパク質を用いた生体内cAMP可視化プローブの開発2016

    • Author(s)
      伊藤幹、原田一貴、Devina Wongso、坪井貴司、北口哲也
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-11-30
    • Related Report
      2016 Research-status Report
  • [Presentation] 蛍光タンパク質を用いた単色型cGMP可視化プローブの開発2016

    • Author(s)
      松田翔吾、原田一貴、伊藤幹、滝澤舞、Devina Wongso、坪井貴司、北口哲也
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-11-30
    • Related Report
      2016 Research-status Report
  • [Patent(Industrial Property Rights)] リガンド蛍光センサータンパク質とその使用2016

    • Inventor(s)
      北口哲也、坪井貴司、上田宏
    • Industrial Property Rights Holder
      北口哲也、坪井貴司、上田宏
    • Industrial Property Rights Type
      特許
    • Filing Date
      2016-05-24
    • Related Report
      2016 Research-status Report
    • Overseas
  • [Patent(Industrial Property Rights)] リガンド蛍光センサータンパク質とその使用2016

    • Inventor(s)
      北口哲也、坪井貴司、上田宏、新井敏
    • Industrial Property Rights Holder
      北口哲也、坪井貴司、上田宏、新井敏
    • Industrial Property Rights Type
      特許
    • Filing Date
      2016-12-02
    • Related Report
      2016 Research-status Report
    • Overseas

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Published: 2016-04-21   Modified: 2022-02-21  

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