辐射诱导RNA分子8-羟基鸟嘌呤动态修饰规律和生物学功能研究

细胞中RNA氧化修饰普遍存在，与多种应激反应和疾病密切相关。8-羟基鸟嘌呤(8-OHG)是RNA氧化损伤修饰的重要形式，但其修饰规律和调控生物分子功能尚未得到阐明。本研究以辐射和过氧化氢诱导RNA分子8-OHG修饰的细胞为模型，采用我们在RNA的m6A修饰建库测序方法基础上改进的HyRIP-Seq和HiCLIP-Seq方法对RNA8-OHG修饰进行高通量分析，获得辐射诱导细胞8-OHG修饰RNA表达谱，揭示其修饰的动态规律；采用全基因组CRISPR-Cas9文库技术筛选参与8-OHG修饰RNA功能调控蛋白分子，同时探讨DNA损伤反应中重要分子ATM(ataxia telangiectasia-mutated gene)在RNA氧化损伤中的作用，阐释8-羟基鸟嘌呤动态修饰RNA的功能和调控作用。本研究将为全面揭示RNA8-OHG动态修饰特征和调控机制奠定基础。

Oxidative modifications of RNAs in cells are ubiquitous and are related to a variety of biological stress responses and diseases. 8-hydroxyguanine is an important modified type of RNA oxidative damage, but its modified pattern and regulatory mechanism have not been elucidated. In this study, the cell model of 8-hydroxyguanidine modification induced by radiation and hydrogen peroxide was used. We improved the HyRIP-Seq and HiCLIP-Seq methods based on the RNA m6A modified library sequencing method for RNA 8-hydroxyguanine modification analysis. The modified pattern and the dynamics of its modification with radiation-induced expression of 8-hydroxyguanine in RNAs will be revealed. Then，whole-genome CRISPR-Cas9 library technology will be used to screen for functional regulatory proteins of 8-hydroxyguanine-modified RNAs and we will investigate the role of ATM (ataxia telangiectasia-mutated gene) in RNA oxidative damage. These results will be disclosure of the characteristics and regulatory mechanisms of dynamic modification of RNAs.