2023 Fiscal Year Final Research Report

Development of pinpoint RNA photo-modification method for mRNA modification and analysis of interstrand crosslinking reaction by photo-catalytic reaction

Research Project

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Project/Area Number	22K14792
Research Category	Grant-in-Aid for Early-Career Scientists
Allocation Type	Multi-year Fund
Review Section	Basic Section 37030:Chemical biology-related
Research Institution	Tohoku University
Principal Investigator	Yamano Yuuhei 東北大学, 多元物質科学研究所, 特任研究員(日本学術振興会特別研究員PD) (50938107)
Project Period (FY)	2022-04-01 – 2024-03-31
Keywords	光修飾 / 光触媒 / 光ラベル化 / 核酸 / 光酸化 / 酸化損傷 / APサイト / 架橋反応
Outline of Final Research Achievements	In this project, we aimed to 1. develop novel photo-modification reaction for nucleic acid, and 2. analyze cross-linking-type nucleic acid lesion under photocatalytic reactions. 1. DNA probes tethered with photocatalysts, Cat-ODNs, are newly designed. Cat-ODN that formed a duplex with the target sequence was exposed to light in the presence of a urazole-type modifier.Intended photochemical modification of nucleic acids was achieved based on the reaction between guanine residues in target sequence and urazole-type modifiers activated by a photocatalyst in Cat-ODN. 2.Confirmed that AP sites were generated in DNA oligomer by irradiating with light in the presence of a photocatalyst.This reaction was mainly mediated by singlet oxygen generated by the photocatalyst. Also suggested that the generated AP sites formed additional cross-linking type lesion with the nucleobases of the complementary strand.
Free Research Field	核酸化学
Academic Significance and Societal Importance of the Research Achievements	１．今回開発した核酸修飾法は、光を照射するだけで標的の配列を修飾できる方法である。従って、選択性と効率に課題は残るものの、長鎖核酸の機能化や蛍光色素によるラベル化といった多岐にわたる応用可能性を秘めていると考えられる。２．本研究では、光架橋型の損傷の発見をきっかけとして、光触媒反応に基づく非酵素的なAPサイト生成を見出した。さらに、その配列・構造依存性や機構についても詳細に検証した。同様の現象に関する詳細な報告はこれまでにない。従って、本研究で得られた知見は未知の損傷修復機構や生命現象の発見・解明にも大いに役立つことが期待され、生物学的にも極めて重要で意義深い試みであると考えられる。