Development of dual-fluorescent ODN probes for SNPs typing

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K05853
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Nihon 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
• Keywords: 蛍光核酸 / プローブ

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.

Free Research Field

生物有機化学

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。