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Establishment of the mothod to quantify methylation efficiency at a specific site

Research Project

Project/Area Number 17K19352
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Research Field Molecular and Genome biology and related fields
Research InstitutionOsaka University

Principal Investigator

Kawahara Yukio  大阪大学, 医学系研究科, 教授 (80542563)

Project Period (FY) 2017-06-30 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
KeywordsRNA修飾 / m6A / メチル化 / エピトランスクリプトーム / ガン / 肥満
Outline of Final Research Achievements

In general, RNAs are subject to post-transcriptional modifications, which affects their fate. Among these modifications, N6-methyladenosine (m6A) is a relatively abundant modification. Given that m6A methylation is related to various diseases, such as obesity and cancer, it is important to know the dynamics of this modification and its regulatory mechanism. However, there has been no sensitive method established to measure methylation efficiency with high accuracy in a site-specific manner. In this study, we found insertion of bridged nucleic acid (BNA) into DNA probes increases the difference in melting temperature between m6A-containing RNA and unmethylated RNA. Applying this principle, we developed a dual BNA probe method, with which we successfully quantified methylation efficiency at m6A sites with high accuracy.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発に成功したBNAプローブ法は、従来定量が不可能であった個別部位でのRNAメチル化効率の測定を可能にした。特に、リボソームRNAなどの豊富に発現しているRNA中のメチル化効率は、高感度で定量できることを示した。本研究の成果は、これまで知ることのできなかったRNAメチル化動態を計測することを可能にしただけでなく、同じ原理を利用すれば他のRNA化学修飾の定量にも応用できることを示唆している。また、その結果としてガンなどの疾患の発症病態の解明に貢献できるものと考える。

Report

(3 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • Research Products

    (15 results)

All 2019 2018 2017 Other

All Int'l Joint Research (2 results) Journal Article (5 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 5 results,  Open Access: 2 results) Presentation (6 results) (of which Int'l Joint Research: 4 results,  Invited: 2 results) Remarks (2 results)

  • [Int'l Joint Research] University of Melbourne(オーストラリア)

    • Related Report
      2018 Annual Research Report
  • [Int'l Joint Research] Institute of Evolutionary Biology/Universitat Pompeu Fabra-CSIC(スペイン)

    • Related Report
      2017 Research-status Report
  • [Journal Article] Quantification of methylation efficiency at a specific N6-methyladenosine position in rRNA by using BNA probes2018

    • Author(s)
      Oshima Takuya、Ishiguro Kensuke、Suzuki Tsutomu、Kawahara Yukio
    • Journal Title

      Chemical Communications

      Volume: 54 Issue: 69 Pages: 9627-9630

    • DOI

      10.1039/c8cc03713b

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] ADAR1-mediated RNA editing is required for thymic self-tolerance and prevention of autoimmunity2018

    • Author(s)
      Taisuke Nakahama, Yuki Kato, Jung In Kim, Tuangtong Vongpipatana, Yutaka Suzuki, Carl Walkley and Yukio Kawahara
    • Journal Title

      EMBO Reports

      Volume: 19 Issue: 12

    • DOI

      10.15252/embr.201846303

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] RNA editing independently occurs at three mir-376a-1 sites and may compromise the stability of the microRNA hairpin.2017

    • Author(s)
      Gallego A, Hartasanchez DA, Braso-Vives M, Garcia-Ramallo E, Lopez-Valenzuela M, Baena N, Guitart M, Fernandez-Bellon H, Kondova I, Bontrop R, Kawahara Y, Espinosa-Parrilla Y
    • Journal Title

      Gene

      Volume: 628 Pages: 109-116

    • DOI

      10.1016/j.gene.2017.07.032

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Matrin3 binds directly to intronic pyrimidine-rich sequences and controls alternative splicing2017

    • Author(s)
      Uemura Y, Oshima T, Yamamoto M, Reyes CJ, Costa Cruz PH, Shibuya T, Kawahara Y
    • Journal Title

      Genes to Cells

      Volume: 22(9) Issue: 9 Pages: 785-798

    • DOI

      10.1111/gtc.12512

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] The RNA-binding protein MARF1 promotes cortical neurogenesis through its RNase activity domain2017

    • Author(s)
      Kanemitsu Y, Fujitani M, Fujita Y, Zhang S, Su YQ, Kawahara Y, Yamashita T
    • Journal Title

      Scientific Reports

      Volume: 7(1) Issue: 1 Pages: 1155-1155

    • DOI

      10.1038/s41598-017-01317-y

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ADAR1-mediated RNA editing is required for thymic selftolerance and inhibition of autoimmunity2019

    • Author(s)
      Nakahama T, Kato Y, Kim JI, Vongpipatana T, Suzuki Y, Walkley CR, Kawahara Y
    • Organizer
      Gordon Research Conference
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ADAR1 regulates early T cell development via MDA5-dependent and -independent pathways2019

    • Author(s)
      Tuangtong Vogpipatana, Taisuke Nakahama, Yuki Kato, Carl Walkley and Yukio Kawahara
    • Organizer
      Gordon Research Conference & Seminar
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Precise determination of responsible ADARs for each RNA editing site in vivo2019

    • Author(s)
      Pedro Herique Costa Cruz, Yuki Kato, Taisuke Nakahama, Toshiharu Shibuya, Yukio Kawahara
    • Organizer
      Gordon Research Conference & Seminar
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The role of ADAR1-mediated RNA editing in development2017

    • Author(s)
      Yukio Kawahara
    • Organizer
      ConBio2017 (2017年度) 生命科学系学会合同年次大会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] The role of RNA editing in T cell development2017

    • Author(s)
      Yukio Kawahara
    • Organizer
      第43回内藤カンファランス
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Matrin3 binds directly to intronic pyrimidine-rich sequences and controls alternative splicing2017

    • Author(s)
      Yuri Uemura, Takuya Oshima1, Munetaka Yamamoto, Charles Jourdan Reyes, Pedro Herique Costa Cruz, Toshiharu Shibuya, Yukio Kawahara
    • Organizer
      RNA2017, The 22nd annual meeting of the RNA society
    • Related Report
      2017 Research-status Report
  • [Remarks]

    • URL

      http://www.med.osaka-u.ac.jp/pub/rna/publications.html

    • Related Report
      2018 Annual Research Report
  • [Remarks] http://www.med.osaka-u.ac.jp/pub/rna/index.html

    • Related Report
      2017 Research-status Report

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Published: 2017-07-21   Modified: 2020-03-30  

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