Development of fluorescent ligands for the analysis of non-coding RNA

2014 Fiscal Year Final Research Report

• Project/Area Number: 24350033
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• 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)
• Co-Investigator(Kenkyū-buntansha): SATO Yusuke 東北大学, 大学院理学研究科, 助教 (90583039)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: ノンコーディングRNA / TAR RNA / microRNA / siRNA / 蛍光性リガンド / スクリーニング / 検出 / イメージング

Outline of Final Research Achievements

In this work, new analytical methods using synthetic fluorescent ligands are successfully developed for the study of non-coding RNA, including trans-activation-responsive region (TAR) RNA of human immunodeficiency virus type 1 (HIV-1), microRNA, and small interfering RNA (siRNA). First, we report on promising binding and signaling functions of thiazole orange (TO) to recognize TAR RNA. We found that TO does bind to selectivity TAR RNA with a nanomolar affinity, and TO was utilized as a fluorescent indicator for the high-throughput screening assay to identify new class of scaffolds of HIV inhibitors. As for the detection of microRNA in vitro, fluorescent ligands are designed and synthesized based on abasic site-binding ligands that are conjugated with cyanine dyes. Furthermore, fluorescent probes are successfully developed based on peptide nucleic acids for intracellular delivery analysis of siRNA.

Free Research Field

分析化学