Development of the chemical reaction probes for intracellular miRNA detection

2023 Fiscal Year Final Research Report

• Project/Area Number: 20K05563
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 34020:Analytical chemistry-related
• Research Institution: Gifu University
• Principal Investigator: Shibata Aya 岐阜大学, 工学部, 助教 (50462693)
• Project Period (FY): 2020-04-01 – 2024-03-31
• Keywords: 遺伝子検出

Outline of Final Research Achievements

One method of nucleic acid sensing in living cells is the chemical reaction probes using a target nucleic acid as a template. The advantage of this probes is that the signal can be amplified by running a reaction cycle using the target nucleic acid as a template. Many of these probes use structural changes in the fluorescent substrate as the key to signal generation. So, it is limiting the wavelengths of fluorescence that can be used. In this study, we aimed to solve this problem by developing new FRET based chemically reaction probe that can be post modified with fluorescent molecules using crick reaction. Herein, we report the development of a molecular transfer probe which triggered by a nucleophilic aromatic substitution reaction. As a result, it was found that the probe detected until 0.25 nM template DNA in the catalytic templated reactions.

Free Research Field

核酸化学

Academic Significance and Societal Importance of the Research Achievements

miRNAは組織または時期特異的に多数のmRNAを制御することで、細胞増殖・アポトーシス・分化などに深く関与している。そのため、生きた細胞内でmiRNAの挙動を調べることは生命現象を理解するうえで重要である。今回、開発した分子移動型遺伝子検出プローブはFRETベースのプローブのため多種多様な蛍光剤を組み合わせることができる。そのため、複数のmiRNAの挙動を同時に観察することが可能になると考えられる。