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Development of dual-fluorescent ODN probes for SNPs typing

Research Project

Project/Area Number 16K05853
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Bio-related chemistry
Research InstitutionNihon University

Principal Investigator

Saito Yoshio  日本大学, 工学部, 准教授 (40385985)

Co-Investigator(Kenkyū-buntansha) 根本 修克  日本大学, 工学部, 教授 (30237812)
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords蛍光核酸 / プローブ / 一塩基多型 / SNPs / 蛍光プローブ / DNA
Outline of Final Research Achievements

The novel environmentally sensitive fluorescent nucleosides, 3,7-bis-(naphthalen-1-ylethynyl)-8-aza-3,7-dideaza-2’-deoxyadenosine (3n7nzA) and 7-(naphthalen-1-ylethynyl)-8-aza-3,7-dideaza-2’-deoxyadenosine (37nzA), have been synthesized. Both 3n7nzA and 37nzA possess large π-conjugated systems which extend into both the major and minor grooves or the major groove alone, respectively. The nucleosides showed large solvatochromic shifts (3n7nzA: Δλ = 45 nm, 37nzA: Δλ = 78 nm) and were examined for their ability to fluorimetrically report hybridization events. When incorporated into DNA probes, the bis-substituted 3n7nzA selectively recognized T base on target strands which was reported by a distinct change in its emission wavelength in the long wavelength region. Thus, 3n7nzA has the potential for use as a fluorescent probe for structural studies of DNAs including the detection of single-base alterations in target DNA sequences.

Academic Significance and Societal Importance of the Research Achievements

極性、粘性やなどの周辺のミクロ環境変化に応じて蛍光強度や波長を鋭敏に変化させる環境感応型蛍光核酸塩基を開発し、標的核酸とハイブリダイズした際のマッチ-ミスマッチの違いによるミクロ環境変化を蛍光色の変化で検出できれば、一塩基変異を含む標的核酸の新たな検出法へと繋がる。さらにこれを応用することで、細胞内での核酸の局所的な構造変化(一塩基変異等も含む)やタンパク質との結合状態変化などを色の変化でリアルタイムに検出できるプローブへと展開が可能である。このような技術は、病院等での瞬時の病気の診断にもつながり、オーダーメイド医療の発展に大きく役立つものと考えられる。

Report

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

    (8 results)

All 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (3 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 3 results,  Acknowledgement Compliant: 1 results) Presentation (4 results) (of which Int'l Joint Research: 2 results,  Invited: 1 results)

  • [Int'l Joint Research] The University of Western Ontario(カナダ)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] A fluorescent 3,7-bis-(naphthalen-1-ylethynylated)-2'-deoxyadenosine analogue reports thymidine in complementary DNA by a large emission Stokes shift2018

    • Author(s)
      M. Yanagi, A. Suzuki, R. H. E. Hudson, Y. Saito
    • Journal Title

      Org. Biomol. Chem.

      Volume: 16 Issue: 9 Pages: 1496-1507

    • DOI

      10.1039/c8ob00062j

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The fluorescently responsive 3-(naphthalen-1-ylethynyl)-3-deaza-2'-deoxyguanosine discriminates cytidine via the DNA minor groove2017

    • Author(s)
      A. Suzuki, M. Yanagi, T. Takeda, R. H. E. Hudson, Y. Saito*
    • Journal Title

      Org. Biomol. Chem.

      Volume: 15 Issue: 37 Pages: 7853-7859

    • DOI

      10.1039/c7ob01931a

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] C2-substituted 8-aza-7-deaza-2'-deoxyadenosines as environmentally sensitive fluorescent nucleosides for discriminating apurinic/apyrimidinic sites in DNA duplex2017

    • Author(s)
      T. Yamauchi, T. Takeda, M. Yanagi, N. Takahashi, A. Suzuki, Y. Saito
    • Journal Title

      Tetrahedron Lett.

      Volume: 58 Issue: 2 Pages: 117-120

    • DOI

      10.1016/j.tetlet.2016.11.029

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] 8-アザ-7-デアザ-2'-デオキシアデノシン誘導体を含む環境感応型蛍光DNAプローブの開発2018

    • Author(s)
      栁昌樹、白岩昭吾、齋藤義雄
    • Organizer
      第29回万有仙台シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] Design and synthesis of C2-substituted 8-aza-7-deaza-2’-deoxyadenosines as environmentally sensitive fluorescent nucleosides2017

    • Author(s)
      Y. Saito, M. Yanagi
    • Organizer
      254th American Chemical Society National Meeting &Exposition
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Design of pyrene-labeled 3-deaza-2’-deoxyadenosines comprising a non-π-conjugated linker: Novel fluorescence quenching-based oligodeoxynucleotide probes2016

    • Author(s)
      Y. Saito, T. Takeda, T. Aso, M. Yanagi, N. Takahashi
    • Organizer
      Asian Photochemistry Conference (APC) 2016
    • Place of Presentation
      シンガポール
    • Year and Date
      2016-12-04
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Synthesis and photophysical properties of environmentally sensitive fluorescent purine nucleosides2016

    • Author(s)
      齋藤義雄
    • Organizer
      平成28年度化学系学協会東北大会
    • Place of Presentation
      いわき明星大学(福島県いわき市)
    • Year and Date
      2016-09-10
    • Related Report
      2016 Research-status Report
    • Invited

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

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