Development of highly sensitive linear probe without stem structure

2015 Fiscal Year Final Research Report

• Project/Area Number: 25248037
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Nagoya University
• Principal Investigator: Asanuma Hiroyuki 名古屋大学, 工学(系)研究科(研究院), 教授 (20282577)
• Co-Investigator(Kenkyū-buntansha): KAMIYA Yukiko 名古屋大学, エコトピア科学研究所, 講師 (00527947) ; KASHIDA Hiromu 名古屋大学, 大学院工学研究科, 准教授 (30452189)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: DNA / PNA / 蛍光プローブ / mRNA / ストランドインベージョン / ペリレン / リニアプローブ

Outline of Final Research Achievements

We have designed new fluorescent probe termed linear probe that does not involve stem structure. In this project, we have successfully developed extremely sensitive linear probe involving both perylenes and anthraquinones as fluorophores and quenchers, respectively; signal-to-back ground ratio became as high as 1600. Due to the remarkable nuclease resistivity, newly designed linear probe could fluorescently detect mRNA in cell. We have further developed strand-invader by combining linear probe and PNA. For this purpose, we multiply introduced ethynylperylenes and anthraquinones into DNA for stable complexation with DNA and suppressed complexation with PNA. When linear probe was incubated double-stranded DNA involving target site in the presence of complementary PNA, we could observe fluorescent increase due to the strand invasion. Even PCR product could be fluorescently labelled in a strand-invading manner.

Free Research Field

核酸有機化学