Development of Micro Arrays for Analyzing Small RNAs

2014 Fiscal Year Final Research Report

• Project/Area Number: 22225003
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Analytical chemistry
• Research Institution: Tohoku University
• Principal Investigator: TERAMAE Norio 東北大学, 理学(系)研究科(研究院), 名誉教授 (70114569)
• Co-Investigator(Kenkyū-buntansha): YAMAGUCHI Akira 茨城大学, 理学部, 准教授 (10359531) ; NISHIZAWA Seiichi 東北大学, 大学院理学研究科, 教授 (40281969) ; SATO Yusuke 東北大学, 大学院理学研究科, 助教 (90583039) ; XU Zhiai 東北大学, 大学院理学研究科, 助教 (50562336)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Keywords: 化学センサー / バイオセンサー / 蛍光性リガンド / 核酸 / ナノ細孔 / 過冷却 / 小分子RNA

Outline of Final Research Achievements

This work aimed to develop new strategy of small RNA analysis based on the RNA-binding fluorescent ligands in confined nanopores. Toward this end, we focused on the design of fluorescent abasic site (AP site)-bidning ligands (APLs) in RNA or DNA/RNA duplexes. We successfully found useful G, C, A, or U-discriminating candidates targeting the duplexes. Moreover, new class of light-up fluorescent ligands was developed based on the conjugation of APLs with cyanine dyes. On the other hand, we revealed several unique features of confined spaces inside the silica and alumina nanopores. Especially, exothermic reactions such as cyclodextrin inclusion and DNA hybridization were found to be accelerated inside the nanopores under supercooled condition. Finally, we found possible applications to detect dozens of attomole nucleic acids and analyze their sequences with single nucleotide resolution by the binding of APL-cyanine dye conjugates inside the confined nanopores under supercooled conditions.

Free Research Field

分析化学