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Development of high-throughput system for extraction of intracellularly localized RNAs

Research Project

Project/Area Number 19K15601
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 34020:Analytical chemistry-related
Research InstitutionKanazawa University

Principal Investigator

ZHOU YUANSHU  金沢大学, ナノ生命科学研究所, 特任助教 (60758556)

Project Period (FY) 2019-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords細胞内回収 / RNA回収 / 走査型イオンコンダクタンス顕微鏡 / RNA局所回収
Outline of Research at the Start

神経細胞の細胞核から樹状突起に輸送された伝令RNAは記憶形成に関与すると期待されている。しかし、標的部位への局所的アクセス方法が難しいことや、解析のスループットの問題などがあり、局在するRNAの機能解析が難しく、未だ証拠となるデータはない。本研究は、多機能ナノピペットを基盤として、RNA輸送の網羅的計測を進める。研究方法としては、ハイスループット的回収システムを開発し、それを海馬神経細胞の樹状突起に存在するRNA内包微粒子の回収に応用する。本研究によってRNA内包微粒子を介した神経細胞RNA輸送の新たな分子機構及び機能が明らかになると期待される。

Outline of Final Research Achievements

RNA transported from the nucleus of a neuron to its dendrites, which divide like branches, are expected to be involved in memory formation. However, functional analysis of localized RNA is difficult due to difficulties in local access methods to target sites and throughput of analysis. In this study, dynamic imaging of neurons and recovery of local RNA were realized by using a multifunctional nanopipette developed by the applicant, which can both measure the non-invasive shape of the cell surface and recover a very small amount of liquid material.

Academic Significance and Societal Importance of the Research Achievements

本研究の手法はナノスケールで生細胞の形状変化を非標識・非接触で取得可能であり、他の技術では計測困難な細胞の局所的変化を可視化できる。また、ナノピペットを探針に用いるため、局所的な薬剤投与や細胞質の回収が可能である。その成果の発展として、RNAとともに回収したタンパク質の質量分析により、RNA輸送のメカニズムと機能の解析が可能になる。さらに、RNAと記憶形成の関連性が解明できれば、RNAの神経細胞間移植による記憶の再現という研究分野の開拓が期待できる。

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (3 results)

All 2021 2020

All Journal Article (3 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 3 results,  Open Access: 3 results)

  • [Journal Article] Electrochemical Quantitative Evaluation of the Surface Charge of a Poly(1-vinylimidazole) Multilayer Film and Application to Nanopore pH Sensor2021

    • Author(s)
      Katsuhiko Sato, Fumiya Sato, Masayuki Kumano, Toshio Kamijo, Takaya Sato, Yuanshu Zhou, Yuri Korchev, Takeshi Fukuma, Tsutomu Fujimura, Yasufumi Takahashi
    • Journal Title

      Electroanalysis

      Volume: 10 Issue: 6 Pages: 121-125

    • DOI

      10.1002/elan.202100041

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Transient Kinetics of O2 Evolution in Photocatalytic Water-Splitting Reaction2020

    • Author(s)
      Kosaka Takumu、Teduka Yuya、Ogura Takuya、Zhou Yuanshu、Hisatomi Takashi、Nishiyama Hiroshi、Domen Kazunari、Takahashi Yasufumi、Onishi Hiroshi
    • Journal Title

      ACS Catalysis

      Volume: 10 Issue: 22 Pages: 13159-13164

    • DOI

      10.1021/acscatal.0c04115

    • NAID

      120006952722

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] High-Speed SICM for the Visualization of Nanoscale Dynamic Structural Changes in Hippocampal Neurons2020

    • Author(s)
      Takahashi Yasufumi、Zhou Yuanshu、Miyamoto Takafumi、Higashi Hiroki、Nakamichi Noritaka、Takeda Yuka、Kato Yukio、Korchev Yuri、Fukuma Takeshi
    • Journal Title

      Analytical Chemistry

      Volume: 92 Issue: 2 Pages: 2159-2167

    • DOI

      10.1021/acs.analchem.9b04775

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research

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Published: 2019-04-18   Modified: 2022-01-27  

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