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Development of construction for Zero Mode Waveguides with low cost and high efficiency and application to analysis of DNA-binding proteins

Research Project

Project/Area Number 16K14669
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Molecular biology
Research InstitutionInstitute of Physical and Chemical Research (2018)
Tokyo Institute of Technology (2017)
Kyoto University (2016)

Principal Investigator

HAN YONG-WOON  国立研究開発法人理化学研究所, 生命医科学研究センター, 研究員 (50566297)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
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)
Keywords1分子イメージング / DNA結合蛋白質 / DNA結合タンパク質 / 1分子イメージング / 生体分子
Outline of Final Research Achievements

The Zero Mode Waveguides (ZMWs) enables us to visualize a single fluorescent molecule under high concentration of the molecules and characterize the protein-protein or protein-DNA complex formation process. I have already established the method for construction of ZMWs and characterized some of DNA-binding proteins using the ZMWs. However, research technique using ZMWs was not still common technique. For availability of ZMWs, in this study, I developed the method for construction of ZMWs using Character Projection electron beam lithography. The cost of ZMWs decreased to 1/5 and the production efficiency was increased to 10-fold compared with my previous construction method for ZMWs.

Academic Significance and Societal Importance of the Research Achievements

生体内の多くの反応は複数種の生体分子が多段階に結合解離を行う一連の過程を経て、進行し、蛍光1分子イメージング技術により、そのような過程をリアルタイムに観察することが可能となった。そして、高濃度で蛍光1分子イメージング観察を可能とするナノ開口基板により、様々な生体分子複合体の結合解離の様子を解析することが可能となった。しかしながら、分子生物学の今後の研究にこの手法を取り入れて行く為には、ナノ開口基板をより安価で効率よく作製する手法を開発する必要があったが、本研究成果により、今後、他の研究者もより簡便にナノ開口基板を利用する研究を開始するできるという点で大きな学術的意義があったと考えている。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (6 results)

All 2018 2017 2016

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

  • [Journal Article] Preferential 5-Methylcytosine Oxidation in the Linker Region of Reconstituted Positioned Nucleosomes by Tet1 Protein2016

    • Author(s)
      Kizaki, Seiichiro; Zou, Tingting; Li, Yue; Han, Yong-Woon; Suzuki, Yuki; Harada, Yoshie; Sugiyama, Hiroshi
    • Journal Title

      Chemistry

      Volume: 22 Issue: 46 Pages: 16598-16601

    • DOI

      10.1002/chem.201602435

    • NAID

      120006460116

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ゼロモード導波路(ZMW)を用いた生体分子複合体の定量分析2018

    • Author(s)
      中尾 公子, 多田隈 尚史, 韓 龍雲, 原田 慶恵
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Holliday構造DNAはRuvA-RuvB複合体形成を促進させる2017

    • Author(s)
      韓 龍雲
    • Organizer
      第40回日本分子生物学会
    • Related Report
      2017 Research-status Report
  • [Presentation] 分光学的手法によるクロマチンリモデリング因子の機能解析2016

    • Author(s)
      韓 龍雲
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県、横浜市)
    • Year and Date
      2016-11-30
    • Related Report
      2016 Research-status Report
  • [Presentation] Characterization of RuvA-RuvB-Holliday junction DNA complex formation using fluorescently labeled RuvA2016

    • Author(s)
      Yong-Woon Han
    • Organizer
      第54回日本生物物理学会年会
    • Place of Presentation
      つくば国際会議場(茨城県、つくば市)
    • Year and Date
      2016-11-25
    • Related Report
      2016 Research-status Report
  • [Presentation] Spectroscopic analysis of fluorescently labeled RuvA for characterization of RuvA-RuvB-Holliday junction DNA complex formation2016

    • Author(s)
      Yong-Woon Han
    • Organizer
      10th 3R Symposium
    • Place of Presentation
      ホテル一畑(島根県、松江市)
    • Year and Date
      2016-11-13
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research

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Published: 2016-04-21   Modified: 2020-03-30  

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