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Development of epitranscriptome analysis method using the specific recognition of antibodies to the higher order structures of RNA

Research Project

Project/Area Number 18K06192
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43050:Genome biology-related
Research InstitutionNational Institute of Advanced Industrial Science and Technology

Principal Investigator

Yoshioka Kyoko  国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (50358321)

Co-Investigator(Kenkyū-buntansha) 栗田 僚二  国立研究開発法人産業技術総合研究所, 生命工学領域, 研究グループ長 (50415676)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
KeywordsRNAエピジェネティクス / N6-メチルアデノシン / イムノアッセイ法 / 表面プラズモン共鳴法 / マイクロデバイス / N6メチルアデノシン / イムノアッセイ / 大腸菌RNA / RNA修飾 / エピジェネティクス / 抗原抗体反応 / トランスクリプトーム
Outline of Final Research Achievements

We developed a method for detecting RNA specific N6-methyladenosine by utilizing the selective recognition of antibodies to a nucleobase in the higher-order structure of the RNA-DNA duplex, especially in the bulge structure, that can move freely. We confirmed that a N6-methyladenosine in a specific sequence of rRNA of E. coli could be selectively detected with our immunoassay method. This immunoassay method was applied to a microdevice using the surface plasmon resonance method, and it was shown that the information of a modified RNA can be rapidly obtained using thi microdevice, with a small amount of RNA sample without the RNA amplification.

Academic Significance and Societal Importance of the Research Achievements

本研究で提案したRNA-DNAハイブリッド鎖高次構造内にある塩基を抗体が選択的に認識できることに基づく検出原理を利用したイムノアッセイ法は、特定のRNA塩基の修飾情報を簡便・迅速に測定できる方法であり、網羅的なエピトランスクリプトーム解析技術とともに、有用な情報が得られる測定法であると考える。将来、特定のRNA修飾と疾病等の関連性が明らかになれば、本イムノアッセイ法を適用したマイクロデバイス等を利用することで、医療現場等で当該RNAの修飾情報をオンサイトで迅速・正確に得ることができると考える。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (23 results)

All 2021 2020 2019 2018

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 4 results) Presentation (17 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Quantitative analysis of global 5-methyl- and 5-hydroxymethylcytosine in TET1 expressed HEK293T cells2020

    • Author(s)
      小島直、須田友美、藤井紳一郎、平野和己、波平昌一、栗田僚二
    • Journal Title

      Biosensors & Bioelectronics

      Volume: 167 Pages: 112472-112478

    • DOI

      10.1016/j.bios.2020.112472

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Microfluidic Sensing System with a Multichannel Surface Plasmon Resonance Chip: Damage-Free Characterization of Cells by Pattern Recognition2020

    • Author(s)
      Sugai Hiroka、Tomita Shunsuke、Ishihara Sayaka、Yoshioka Kyoko、Kurita Ryoji
    • Journal Title

      Analytical Chemistry

      Volume: 92 Issue: 22 Pages: 14939-14946

    • DOI

      10.1021/acs.analchem.0c02220

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Sequential Assessment of Multiple Epigenetic Modifications of Cytosine in Whole Genomic DNA by Surface Plasmon Resonance2019

    • Author(s)
      栗之丸隆章、小島直、栗田僚二
    • Journal Title

      Analytical Chemistry

      Volume: 91 Issue: 21 Pages: 13933-13939

    • DOI

      10.1021/acs.analchem.9b03423

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Immobilization of DNA on Biosensing Devices with Nitrogen Mustard Modified Linkers2019

    • Author(s)
      栗之丸隆章、小島直、栗田僚二
    • Journal Title

      Current Protocols in Nucleic Acid Chemistry

      Volume: 77 Issue: 1

    • DOI

      10.1002/cpnc.85

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Immobilization of DNA with nitrogen mustard-biotin conjugate for global epigenetic analysis2018

    • Author(s)
      小島直、須田友美、栗之丸隆章、栗田僚二
    • Journal Title

      Analytica Chimica Acta

      Volume: 1043 Pages: 107-114

    • DOI

      10.1016/j.aca.2018.09.008

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] N6-Methylation Assessment in Escherichia coli 23S rRNA Utilizing a Bulge Loop in an RNA-DNA Hybrid2018

    • Author(s)
      吉岡恭子、栗田僚二
    • Journal Title

      Analytical Chemistry

      Volume: 90 Issue: 12 Pages: 7578-7582

    • DOI

      10.1021/acs.analchem.8b01223

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] 表面プラズモン共鳴の応答パターンを出力可能なマイクロ流体デバイスによる細胞評価2021

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      2020年度細胞アッセイ研究会シンポジウム
    • Related Report
      2020 Annual Research Report
  • [Presentation] パターン認識に基づいて細胞を評価するセンサチップの開発2021

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      SATテクノロジー・ショーケース 2021
    • Related Report
      2020 Annual Research Report
  • [Presentation] 表面プラズモン共鳴の特徴パターンを出力するマルチチャネル型チップによる培養細胞の非破壊的評価2020

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      第80回分析化学討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Immunochemical Assessment of 5-Methylcytosine and 5-Hydroxymethylcytosine Using Nitrogen Mustard Modified Linkers2019

    • Author(s)
      小島直、須田友美、栗之丸隆章、栗田僚二
    • Organizer
      The 46th International Symposium on Nucleic Acids Chemistry, The 3rd Annual Meeting of Japan Society of Nucleic Acids Chemistry
    • Related Report
      2019 Research-status Report
  • [Presentation] Development of New Material and Device for Biomolecular Analysis2019

    • Author(s)
      栗田僚二
    • Organizer
      4th International Conference on Nutraceuticals and Chronic Diseases
    • Related Report
      2019 Research-status Report
  • [Presentation] エピゲノムの迅速分析に向けた材料創成とマイクロデバイス化2019

    • Author(s)
      栗田僚二
    • Organizer
      日本分析化学会第68年会
    • Related Report
      2019 Research-status Report
  • [Presentation] Electrochemiluminescence derived from Surface Accumulable Co-reactant for Biomolecular Determinations2019

    • Author(s)
      栗田僚二
    • Organizer
      17th ISEAC & 3rd ECL
    • Related Report
      2019 Research-status Report
  • [Presentation] 特異/非特異プローブによるマイクロフルイディクスでの生体分子検出2019

    • Author(s)
      栗田僚二
    • Organizer
      JASISカンファレンス
    • Related Report
      2019 Research-status Report
  • [Presentation] 表面プラズモン共鳴チップによるパターン認識に基づく培養細胞の非破壊的評価2019

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      第14回ナノ・バイオメディカル学会
    • Related Report
      2019 Research-status Report
  • [Presentation] 生体試料の特徴パターンを出力する表面プラズモン共鳴センサチップの開発2019

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      化学とマイクロ・ナノシステム学会第39回研究会
    • Related Report
      2019 Research-status Report
  • [Presentation] 細胞の特徴パターンを出力する表面プラズモン共鳴センサの開発2019

    • Author(s)
      菅井祥加、冨田峻介、石原紗綾夏、吉岡恭子、栗田僚二
    • Organizer
      Chem-Bio Joint Seminar 2019
    • Related Report
      2019 Research-status Report
  • [Presentation] 抗体のバルジ構造特異的な認識を利用したrRNA修飾塩基検出法2018

    • Author(s)
      吉岡恭子、栗田僚二
    • Organizer
      日本分析化学会第67年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Detection of Cytosine Variants of DNA Using Surface Plasmon Resonance-Based Immunoassay2018

    • Author(s)
      栗之丸隆章、小島直、栗田僚二
    • Organizer
      第45回国際核酸化学シンポジウム日本核酸化学会第2回年会
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] エピゲノム修飾の包括的評価にむけたSPRイムノアッセイの開発2018

    • Author(s)
      栗之丸隆章、小島直、栗田僚二
    • Organizer
      第12回バイオ関連化学シンポジウム
    • Related Report
      2018 Research-status Report
  • [Presentation] 核酸固定化試薬の開発とメチル化シトシンイムノアッセイへの応用2018

    • Author(s)
      小島直、栗之丸隆章、須田友美、栗田僚二
    • Organizer
      日本核酸医薬学会第4回年会
    • Related Report
      2018 Research-status Report
  • [Presentation] シトシンバリアントの一括計測に向けたSPRイムノアッセイの開発2018

    • Author(s)
      栗之丸隆章、小島直、栗田僚二
    • Organizer
      化学とマイクロ・ナノシステム学会第37回研究会
    • Related Report
      2018 Research-status Report
  • [Presentation] Immobilization of Genomic DNA with Bifunctional Linker Molecules for 5-Methylcytosine Immunoassay2018

    • Author(s)
      小島直、栗之丸隆章、須田友美、栗田僚二
    • Organizer
      9th Annual symposium of the Indian Scientists Association in Japan
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research

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Published: 2018-04-23   Modified: 2022-01-27  

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