Development of fluorescent small probes for RNA imaging in cells

2014 Fiscal Year Final Research Report

• Project/Area Number: 25620102
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Analytical chemistry
• Research Institution: Tohoku University
• Principal Investigator: NISHIZAWA Seiichi 東北大学, 理学(系)研究科(研究院), 教授 (40281969)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: siRNA / 蛍光プローブ / イメージング

Outline of Final Research Achievements

In this work, synthetic fluorescent probes are successfully developed for intracellular delivery analysis of small interfering RNAs (siRNAs). The probes are based on peptide nucleic acids that are conjugated with thiazole orange (TO) and pyrene to their C- and N-termini, respectively. The probe is able to selectively bind to 3’-overhanging two nucleotides uniquely found in siRNAs, and the binding is accompanied by a significant light-up response due to intercalation of the TO moiety into the double-stranded regions near overhangs. In contrast to typical methods using fluorophore-labeled siRNA, the present probe does not compromise the siRNA silencing activity, and is applicable to the analysis of both cellular uptake and the intracellular disassembly of the polyplexes. The use of the probe thus allows a more detailed and quantitative analysis of the siRNA delivery process, which would be very useful for siRNA-related study.

Free Research Field

分析化学