Development of the method for large-scale analysis of the reactive molecules to RNA higher-ordered structure

2022 Fiscal Year Final Research Report

• Project/Area Number: 20H02855
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Tohoku University
• Principal Investigator: Nagatsugi Fumi 東北大学, 多元物質科学研究所, 教授 (90208025)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: RNA高次構造 / 選択的アルキル化 / 大規模解析 / RNA標的創薬

Outline of Final Research Achievements

Since non-coding RNAs (ncRNAs) form many similar higher-order structures, there are limited reports of RNA-binding small molecules targeting them. In this study, we aimed to establish a method for analyzing the true selectivity of chemically reactive molecules for various RNA higher-order structures and to apply this method to new structure-reactivity relationship studies. As a result, using vinylquinazolinone derivatives (VQ), which are OFF-ON type reactive molecules that induce alkylation reactions in close proximity to the target structure, we succeeded in obtaining sequence information on RNAs that undergo efficient alkylation by barcode microarray method. Furthermore, it is found that VQ derivative can effectively alkylate to pre-miR221, which was obtained as a high-ranking sequence. Pre-miR221 is overexpressed in cancer stem cells of colorectal cancer and breast cancer, and is of interest as a cancer therapeutic target

Free Research Field

核酸化学、ケミカルバイオロジー

Academic Significance and Societal Importance of the Research Achievements

本研究では標的構造に近接しアルキル化反応が誘起されるOFF-ON型反応性分子を用いて、バーコードマイクロアレイ法により効率的にアルキル化が進行するRNAの大規模配列情報を得ることに成功した。今回、確立した解析手法はPCRを必要とせず、反応性分子に適用可能であり非常に独自性が高い。さらに、本解析法では10万種類以上の配列を含む多様な高次構造を持つRNAライブラリから、合成した低分子が反応する配列を簡便にランク化可能である。合成した反応性分子ごとに反応嗜好性のビッグデータを得られることから、将来的にはRNA標的AI創薬の重要な技術となると期待され社会的な意義も非常に高いと考えられる。