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Development of a method for spatio-temporal analysis of gene expression based on RNA visualization and bioluminescence imaging techniques

Research Project

Project/Area Number 19H02745
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 34020:Analytical chemistry-related
Research InstitutionThe University of Tokyo

Principal Investigator

Yoshimura Hideaki  東京大学, 大学院理学系研究科(理学部), 助教 (90464205)

Project Period (FY) 2019-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2021: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
Keywordsバイオイメージング / 生物発光 / RNA / 細胞 / 遺伝子発現 / 顕微鏡
Outline of Research at the Start

本研究は申請者が独自に構築したRNAイメージング技術を基盤として、多数細胞からなる集団において1細胞ごとの生細胞内遺伝子発現の時空間変化を、細胞内局在を解析可能な空間解像度で数日以上にわたって定量分析する、世界初の技術創出を目指す。この技術を用いて、多細胞集団の中で個々の細胞に確率論的に生じる細胞生理現象、すなわちリプログラミング(iPS細胞化)や分化誘導、ガン化などの細胞現象が生じるための必要十分条件となる遺伝子発現時空間特性を同定する。さらに本手法の原理実証のため、体細胞のリプログラミングなどの細胞生理現象について本手法を適用し、現象を生じる遺伝子発現時空間特性の必要十分条件を検討する。

Outline of Final Research Achievements

The present research aims to develop an RNA probe using bioluminescence to monitor time-course localization alteration of the endogenous target RNA in living cells. In an experiment of the probe solution, the probe quickly responds to the addition and degradation of the target RNA reversibly. Then, the probe expression plasmid was induced into living mammalian cells, and bioluminescence imaging of the cells was observed. In this observation, the target RNA, beta-actin mRNA, formed granule-like structures in the cells, to which growth factor stimulation was administered. The granule-like structures were then concentrated in the expanding cell edge region in response to the administrated stimulation. These results demonstrated the performance of the present probe in monitoring the RNA localization alteration accompanied by cellular physiological events.

Academic Significance and Societal Importance of the Research Achievements

本研究の結果から、開発したプローブは生細胞内において標的とする内在性RNAの細胞現象に伴う局在変化を可視化追跡可能な性能を有することが示された。本プローブの特徴として、標的とするRNAの配列に応じて、そのRNAを標的とするようにカスタムメイドにデザインできることがある。すなわち、本プローブの原理を用いて様々なRNAを標的として生細胞内で局在変化を経時観察できるようになる。またRNAは遺伝子発現産物であり、各種網羅的解析により同定されたRNAが標的とする生理機能の発現過程において示す動態解析にも応用可能である。

Report

(4 results)
  • 2022 Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (16 results)

All 2022 2021 2020 2019

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 1 results) Presentation (6 results) (of which Int'l Joint Research: 1 results,  Invited: 5 results) Book (4 results)

  • [Journal Article] A Series of Furimazine Derivatives for Sustained Live-Cell Bioluminescence Imaging and Application to the Monitoring of Myogenesis at the Single-Cell Level2022

    • Author(s)
      Orioka Mariko、Eguchi Masatoshi、Mizui Yuki、Ikeda Yuma、Sakama Akihiro、Li Qiaojing、Yoshimura Hideaki、Ozawa Takeaki、Citterio Daniel、Hiruta Yuki
    • Journal Title

      Bioconjugate Chemistry

      Volume: 33 Issue: 3 Pages: 496-504

    • DOI

      10.1021/acs.bioconjchem.2c00035

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Triple-color single-molecule imaging for analysis of the role of receptor oligomers in signal transduction2022

    • Author(s)
      Yoshimura Hideaki
    • Journal Title

      Biophysics and Physicobiology

      Volume: 19 Issue: 0 Pages: n/a

    • DOI

      10.2142/biophysico.bppb-v19.0007

    • ISSN
      2189-4779
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Discovery of a phase-separating small molecule that selectively sequesters tubulin in cells2022

    • Author(s)
      Ado Genyir、Noda Naotaka、Vu Hue T.、Perron Amelie、Mahapatra Amarjyoti D.、Arista Karla Pineda、Yoshimura Hideaki、Packwood Daniel M.、Ishidate Fumiyoshi、Sato Shin-ichi、Ozawa Takeaki、Uesugi Motonari
    • Journal Title

      Chemical Science

      Volume: 13 Issue: 19 Pages: 5760-5766

    • DOI

      10.1039/d1sc07151c

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Long-term single cell bioluminescence imaging with C-3 position protected coelenterazine analogues2021

    • Author(s)
      Mizui Yuki、Eguchi Masatoshi、Tanaka Masanobu、Ikeda Yuma、Yoshimura Hideaki、Ozawa Takeaki、Citterio Daniel、Hiruta Yuki
    • Journal Title

      Organic & Biomolecular Chemistry

      Volume: 19 Issue: 3 Pages: 579-586

    • DOI

      10.1039/d0ob02020f

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Signaling activations through G-protein-coupled-receptor aggragations.2020

    • Author(s)
      Watabe, M., Yoshimura, H., Ajunan, S.N.V., Kaizu, K., Takahashi, K.
    • Journal Title

      Phys. Rev. E

      Volume: 102 Issue: 3 Pages: 032413-032413

    • DOI

      10.1103/physreve.102.032413

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Synergetic roles of Formyl Peptide Receptor 1 oligomerization in ligand-induced signal transduction2020

    • Author(s)
      Nishiguchi T, Yoshimura H, Kasai RS, Fujiwara TK, Ozawa T
    • Journal Title

      ACS Chem. Biol.

      Volume: 15 Issue: 9 Pages: 2577-2587

    • DOI

      10.1021/acschembio.0c00631

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Presentation] 生細胞1分子観察による分子集合体形成とシグナル入力反応の可視化分析2021

    • Author(s)
      吉村英哲、西口知輝、小澤岳昌
    • Organizer
      日本分析化学会第70年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 蛍光イメージング分光法 生細胞への応用と蛍光ライブイメージング2021

    • Author(s)
      吉村英哲
    • Organizer
      光とレーザーの科学技術フェア2021「分光フェア」分光セミナー
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Oligomer Formation and Signal Transduction of GPCR - A Single-Molecule Live Imaging Study -2021

    • Author(s)
      Hideaki Yoshimura
    • Organizer
      14th International Symposium on Nanomedicine joint with 11th Taiwan-Japan-Korea Nanomedicine Meeting
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] 生命の仕組みを解明する蛍光分析 -生きたままの細胞を対象に-2020

    • Author(s)
      吉村英哲
    • Organizer
      光とレーザーの科学技術フェア2020
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] The roles of receptor oligomerization for signal transduction - A study through single-molecule live-cell imaging-2019

    • Author(s)
      Hideaki Yoshimura
    • Organizer
      13th International Symposium on Nanomedicine
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 分子動態からメカニズムを探る -生細胞1分子イメージングを用いたアプローチ-2019

    • Author(s)
      吉村英哲
    • Organizer
      第11回光塾
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Book] Optical monitoring of single molecule dynamics of RNA in living cells2021

    • Author(s)
      Hideaki Yoshimura*, Takeaki Ozawa
    • Total Pages
      12
    • Publisher
      Springer Series in Chemical Physics,”Progress in Photon Science”
    • Related Report
      2021 Annual Research Report
  • [Book] 日本分光学会監修 『紫外可視・蛍光分光法』 第3章 蛍光イメージング分光法(FRET, FCS, FCCS):生細胞への応用2021

    • Author(s)
      吉村英哲
    • Total Pages
      14
    • Publisher
      講談社サイエンティフィク刊
    • Related Report
      2021 Annual Research Report
  • [Book] 日本分光学会監修 『紫外可視・蛍光分光法』 付録 蛍光プローブに用いられる色素の一覧2021

    • Author(s)
      吉村英哲
    • Total Pages
      16
    • Publisher
      講談社サイエンティフィク刊
    • Related Report
      2021 Annual Research Report
  • [Book] 発光イメージング実験ガイド プロトコール編 II-4 「二分割ルシフェラーゼ再構成法による発光プローブ」2019

    • Author(s)
      永井 健治、小澤 岳昌
    • Total Pages
      223
    • Publisher
      羊土社
    • ISBN
      9784758122405
    • Related Report
      2019 Annual Research Report

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Published: 2019-04-18   Modified: 2024-01-30  

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