Real-Time Single-Molecule Observation of DNA Conformational Cahnges

• Project/Area Number: 17H03088
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Osaka University
• Principal Investigator: KAWAI KIYOHIKO 大阪大学, 産業科学研究所, 准教授 (50314422)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2017: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: DNA / 高次構造 / 1分子検出 / 構造転移 / 速度 / 蛍光 / blinking / １分子計測 / 1分子計測 / DNA高次構造 / 蛍光相関分光

Outline of Final Research Achievements

We have developed a single-molecule analysis and diagnosis method based on the understanding, control, and utilization of fluorescence blinking (KACB method).
By controlling the blinking by oxidation/reduction reactions, we achieved single molecule discrimination of DNA hairpin, double-stranded, B-type, and A-type structures, and developed a new method for single molecule detection of DNA.
By tracking the temporal change of blinking pattern, we succeeded in single molecule detection of structural transition of functional RNA.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した、KACB法は、様々な分子の1分子検出、そして、その分子のダイナミクスに関する情報の抽出を可能年、様々な、1分子分析、診断法の開発へとつながる基盤技術として期待される。2014年にノーベル化学賞が贈られた、超解像顕微鏡の開発においては、1つ1つ蛍光分子から点を描き、点描画のように高解像度の画像が得られる。本研究では、蛍光分子から点を描くだけでなく、各蛍光分子から発せられる情報をもマッピング可能な技術へと発展すると期待される。

Research Products

• [Int'l Joint Research] imec(ベルギー)
• [Int'l Joint Research] ETH Zurich/ University of Zurich(スイス)
• [Journal Article] Single-Molecule Study of Redox Reaction Kinetics by Observing Fluorescence Blinking (2021)
  • Author(s): Kawai Kiyohiko、Fujitsuka Mamoru、Maruyama Atsushi
  • Journal Title: Accounts of Chemical Research Volume: 54 Issue: 4 Pages: 1001-1010
  • DOI: 10.1021/acs.accounts.0c00754
  • Peer Reviewed
• [Journal Article] Dynamics of Single‐Stranded RNA Looping Probed and Photoregulated by Sulfonated Pyrene (2020)
  • Author(s): J. Xu, S. Tojo, M. Fujitsuka and K. Kawai
  • Journal Title: ChemistrySelect Volume: 5 Issue: 26 Pages: 8002-8008
  • DOI: 10.1002/slct.202002231
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Kinetics of Photoinduced Reactions at the Single‐Molecule Level: The KACB Method (2020)
  • Author(s): Kawai Kiyohiko、Maruyama Atsushi
  • Journal Title: Chemistry - A European Journal Volume: 26 Issue: 35 Pages: 7740-7746
  • DOI: 10.1002/chem.202000439
  • Peer Reviewed
• [Journal Article] Sulfonated Pyrene as a Photoregulator for Single-Stranded DNA Looping (2020)
  • Author(s): Jie Xu, Shunichi Miyamoto, Sachiko Tojo and Kiyohiko Kawai
  • Journal Title: Chemistry - A European Journal Volume: 26 Issue: 22 Pages: 5075-5084
  • DOI: 10.1002/chem.202000184
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Single-Molecule Monitoring of the Structural Switching Dynamics of Nucleic Acids through Controlling Fluorescence Blinking (2017)
  • Author(s): Kawai Kiyohiko、Miyata Takafumi、Shimada Naohiko、Ito Syoji、Miyasaka Hiroshi、Maruyama Atsushi
  • Journal Title: Angew. Chem. Int. Ed. Volume: 56 Issue: 48 Pages: 15329-15333
  • DOI: 10.1002/anie.201708705
  • Peer Reviewed / Open Access
• [Presentation] Sulfonated Pyrene as Both a Probe and a Photoregulator for Single-Stranded Oligonucleotide Looping (2020)
  • Author(s): Jie, Xu; Shunichi, Miyamoto; Sachiko, Tojo; Mamoru Fujitsuka; Kiyohiko Kawai
  • Organizer: 光化学討論会2020
• [Presentation] 核酸1分子を見つける、調べる (2019)
  • Author(s): 川井 清彦
  • Organizer: 第22回生命化学研究会
  • Invited
• [Presentation] 蛍光blinking制御によるRiboswitch構造変化の1分子観測 (2019)
  • Author(s): 川井 清彦
  • Organizer: 日本分析化学会第68回年会
  • Invited
• [Presentation] Single-Molecule Level Monitoring of Nucleic Acids Conformational Changes by Controlling the Fluorescence Blinking (2019)
  • Author(s): Kiyohiko Kawai
  • Organizer: π-System Figuration European-Japanese Workshop
  • Int'l Joint Research / Invited
• [Presentation] Single-Molecule Level Analysis of Nucleic Acids Structure by Controlling the Blinking (2019)
  • Author(s): Kiyohiko Kawai, Takafumi Miyata, Naohiko Shimada, Atsushi Maruyama
  • Organizer: ISNAC 2019
  • Int'l Joint Research
• [Presentation] rKACB法による核酸構造の1分子分析 (2019)
  • Author(s): 川井 清彦、宮田 貴史、嶋田 直彦、丸山 厚
  • Organizer: 第13回バイオ関連化学シンポジウム2019
• [Presentation] 蛍光のblinking観測による核酸構造の1分子分析 (2018)
  • Author(s): 川井清彦
  • Organizer: 日本化学会 第98春季年会 (2018)
  • Int'l Joint Research / Invited
• [Presentation] DNA STRUCTURAL CHANGES MONITORED BY CONTROLLING THE FLUORESCENCE BLINKING (2017)
  • Author(s): 川井 清彦、丸山 厚、真嶋 哲朗
  • Organizer: The 15th Conference on Methods and Applications in Fluorescence
  • Int'l Joint Research
• [Book] 核酸科学ハンドブック (2020)
  • Author(s): 日本核酸化学会、杉本 直己
  • Total Pages: 576
  • Publisher: 講談社
  • ISBN: 9784065207864
• [Remarks] 研究紹介： 現象を調べ、わかったことを積極的に活用することを目指しています！
  • URL: https://www.sanken.osaka-u.ac.jp/labs/mec/kawairesearch.html
• [Remarks] Single-molecule level analysis
  • URL: https://www.sanken.osaka-u.ac.jp/labs/mec/kawairesearch-e.html