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Photon Upconversion Fluorescent Probe for Sequence Selective Detection/Imagine of Double-Stranded DNA

Research Project

Project/Area Number 20H02768
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 34020:Analytical chemistry-related
Research InstitutionKyushu University

Principal Investigator

Nakano Koji  九州大学, 工学研究院, 准教授 (10180324)

Co-Investigator(Kenkyū-buntansha) 石松 亮一  九州大学, 工学研究院, 助教 (90512781)
Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000)
Fiscal Year 2022: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2021: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2020: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Keywords蛍光プローブ / フォトンアップコンバージョン / 二重鎖DNA / 分子イメージング / エピジェネティック
Outline of Research at the Start

試料ダメージが少ない長波長の励起光でも観察できるフォトンアップ コンバージョン蛍光を組み込んだDNAプローブの研究を行う。DNA結合性部位には二重らせん小溝に結合するピロール・イミダゾール・ポリアミド、またはペプチド核酸を基本骨格に選ぶ。アップコンバージョン発光に必要な蛍光性置換基の組込み、DNAへの結合に伴ってコンホメーションが変化して発光をON-OFF制御するデザインにより目的の化合物を合成する。DNAとの結合反応や発光特性を詳しく調べ、培養細胞に導入したプラスミド、さらにはゲノムDNAのin vivoイメージングの研究を通じてゲノム研究のための新しいアプリケーションを提供する。

Outline of Final Research Achievements

We report a new fluorescent probe that utilizes triplet-triplet annihilation-based photon upconversion. In particular, for analyses of the androgen-responsive sequence (ARE) in genomic DNA, we designed a type of ARE-binding pyrrole-imidazole polyamides incorporating Pt-coproporphyrin (PtCP) and anthracene (ANTH). We successfully developed a practical synthetic route in the first half of the research period. Subsequently, we studied the binding behavior to the ARE fragment and measured the fluorescence spectra for the ARE-DNA probe using a laser excitation fluorescence spectrophotometer. At last, we found that when the PtCP in the molecule was excited with excitation light at 533 nm, the molecule gave the phosphorescence of PtCP (650 nm) and the fluorescence from ANTH (480 nm). Therefore we concluded that, for the first time, we successfully developed a photon upconversion probe for DNA analyses.

Academic Significance and Societal Importance of the Research Achievements

基本的な励起一重項からの発光をはじめ蛍光共鳴エネルギー移動、遅延発光を利用する方法など、蛍光プローブは細胞のin vivo観察で成果を収めてきた。しかし、可視領域の蛍光イメージを得るためには強力な紫外光励起が必要で、細胞は深刻なダメージを受けてしまう。このような状況のもと、低損傷な長波長の可視光で励起でき、より短波長側で蛍光を示すDNA結合性蛍光プローブを開発した。これは世界で初めての例であり学術的な意義は大きい。また、二光子励起蛍光法のように取扱いが難しく、大掛かりで高価な装置は必要ないので、実用化の後には、ベンチトップ観察法として細胞研究や医療診断に貢献できる点で社会的な意義は大きい。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (20 results)

All 2022 2021 2020 Other

All Journal Article (9 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 9 results,  Open Access: 2 results) Presentation (10 results) (of which Int'l Joint Research: 2 results) Remarks (1 results)

  • [Journal Article] Development of IgY-Based Indirect Competitive ELISA for the Detection of Fluoroquinolone Residues in Chicken and Pork Samples2022

    • Author(s)
      Sumed Yadoung, Ryoichi Ishimatsu, Zhen-Lin Xu, Korawan Sringarm, Supansa Pata, Marninphan Thongkham, Somporn Chantara, Mookda Pattarawarapan, Surat Hongsibsong
    • Journal Title

      Antibiotics

      Volume: 11 Issue: 11 Pages: 1512-1512

    • DOI

      10.3390/antibiotics11111512

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Development of a facile time-resolved spectroelectrochemical method: An application to determine the rate constant of protonation for anions of 9,10-diphenylanthracene, biphenyl, and p-quaterphenyl2022

    • Author(s)
      Ryoichi Ishimatsu, Yoshie Furukawa, Koji Nakano,
    • Journal Title

      Journal of Electroanalytical Chemistry

      Volume: 921 Pages: 116684-116684

    • DOI

      10.1016/j.jelechem.2022.116684

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sky-blue electrogenerated chemiluminescence using anthracene derivatives as host and guest molecules2022

    • Author(s)
      Emiri Kato, Ryoichi Ishimatsu, Yugo Koinuma, Jun Mizuno, Takashi Kasahara
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 61 Issue: 6 Pages: 060903-060903

    • DOI

      10.35848/1347-4065/ac671a

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Microfluidic electrogenerated chemiluminescence cells using aluminum-doped zinc oxide nanoparticles as an electron injection layer2022

    • Author(s)
      Okada Koji、Ishimatsu Ryoichi、Mizuno Jun、Kasahara Takashi
    • Journal Title

      Sensors and Actuators A: Physical

      Volume: 334 Pages: 113329-113329

    • DOI

      10.1016/j.sna.2021.113329

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Dynamic control of reflective/diffusive optical surfaces on EGaIn liquid metal2021

    • Author(s)
      Nakakubo Keisuke、Yoshioka Hiroaki、Morita Kinichi、Ishimatsu Ryoichi、Kiani Abolfazl、Hallen Hans、Dickey Michael D.、Oki Yuji
    • Journal Title

      Optical Materials Express

      Volume: 11 Issue: 7 Pages: 2099-2099

    • DOI

      10.1364/ome.425432

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Green electrogenerated chemiluminescence using a quinacridone derivative as a guest molecule2021

    • Author(s)
      Koinuma Yugo、Ishimatsu Ryoichi、Kato Emiri、Mizuno Jun、Kasahara Takashi
    • Journal Title

      Electrochemistry Communications

      Volume: 127 Pages: 107047-107047

    • DOI

      10.1016/j.elecom.2021.107047

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Direct Electron Transfer Reaction of Cytochrome c Immobilized on a Bare ITO Electrode2021

    • Author(s)
      Matsuda Naoki、Okabe Hirotaka、Nagamura Toshihiko、Nakano Koji
    • Journal Title

      Bulletin of the Chemical Society of Japan

      Volume: 94 Issue: 2 Pages: 433-439

    • DOI

      10.1246/bcsj.20200192

    • NAID

      130007995392

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Covalent Hyperbranched Polymer Self-Assemblies of Three-Way Junction DNA for Single-Molecule Devices22020

    • Author(s)
      Koji Nakano, Takafumi Sawada, Yoshifumi Mori, Kohei Morita, Ryoichi Ishimatsu
    • Journal Title

      Langmuir

      Volume: 91 Issue: 34 Pages: 10166-10174

    • DOI

      10.1021/acs.langmuir.0c01621

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enzyme-linked immunosorbent assay based on light absorption of enzymatically generated aniline oligomer: Flow injection analysis for 3-phenoxybenzoic acid with anti-3-phenoxybenzoic acid monoclonal antibody2020

    • Author(s)
      Ryoichi Ishimatsu, Shinichi Shimizu, Surat Hongsibsong, Koji Nakano, Chacriya Malasuk, Yuji Oki, Kinichi Morita
    • Journal Title

      Talanta

      Volume: 218 Pages: 121102-121102

    • DOI

      10.1016/j.talanta.2020.121102

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] いくつかの芳香族炭化水素化合物のラジカルアニオンに対するプロトン付加反応速度:分光電気化学的測定2022

    • Author(s)
      石松 亮一、中野 幸二
    • Organizer
      第82回分析化学討論会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 遺伝子のアップコンバージョン蛍光分析のためのドナー/アクセプター修飾ピロール-イミダゾールポリアミドの研究2022

    • Author(s)
      花山 佳祐、中野 幸二、石松 亮一
    • Organizer
      日本分析化学会第71年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 電解生成したラジカルアニオンによる PTFE の表面処理法の検討2022

    • Author(s)
      石松 亮一、中野 幸二
    • Organizer
      日本分析化学会第71年会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Speciality peptides for sub-macromolecular probe for chemical analysis: D/A-modified pyrrole-imidazole polyamide for photon upconversion DNA analysis2022

    • Author(s)
      Keisuke Hanayama, Ryoichi Ishimatsu, Koji NAKANO
    • Organizer
      The 3rd Kyushu-Mainz Chemistry Symposium & The 7th CMS International Symposium (CMS-7)
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] マイクロペルオキシダーゼ/バニロイドレセプターハイブリッド15量体ペプチド酵素の化学合成とペルオキシダーゼ反応の基礎検討2021

    • Author(s)
      中野幸二,木下美里,姫野俊基,石松亮一
    • Organizer
      第81回分析化学討論会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ペプチド核酸/タンパク質酵素ハイブリット分子-PNAzymeの合成とバイオ分析への応用2021

    • Author(s)
      小林 諒祐,中野 幸二,石松 亮一
    • Organizer
      日本分析化学会第70回年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 分光電気化学測定法によるピレンのラジカルアニオンダイマーの検出2021

    • Author(s)
      石松 亮一,中野 幸二
    • Organizer
      日本分析化学会第70回年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] PNAzyme: Monolithic peptide nucleic acid peptide sequence hybrid functioning as molecular recognition catalyst element for nucleic acids analysis2021

    • Author(s)
      K. Nakano, J. Tanabe, Y. Sezaki, R. Kobayashi, R. Ishimatsu
    • Organizer
      Pacifichem 2021
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ペルオキシダーゼ活性を持つペプチド核酸モノリスハイブリッドーPNAzymeの合成と遺伝子センシング応用2020

    • Author(s)
      中野幸二,田邊潤壱,石松亮一
    • Organizer
      日本分析化学会第70年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ピレスロイド系殺虫剤の体内代謝物である3-phenoxybenzoic acid検出に向けたフローインジェクションELISA法の開発2020

    • Author(s)
      石松亮一,清水慎一,中野幸二
    • Organizer
      日本分析化学会第70年会
    • Related Report
      2020 Annual Research Report
  • [Remarks] 九州大学大学院工学研究院応用化学部門 化学環境工学講座 研究紹介

    • URL

      http://www.cstf.kyushu-u.ac.jp/~chemenvironlab/detail.html

    • Related Report
      2020 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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