N6-异戊烯基腺苷的化学修饰和标记

本申请项目在国内外RNA表观遗传修饰研究的基础上，针对tRNA中i6A这种具有重要生物学功能的化学修饰开展高效、高选择性和高灵敏度的化学调控与标记的研究。将突破传统的基因敲除/蛋白过表达的传统研究思路，利用腺苷碱基N6位上的异戊烯基具有较高活性的特性，发展结构可控的化学氧化去修饰体系和试剂，通过选择性地下调细胞内i6A的含量，研究其表达数量与细胞表型之前的联系。通过筛选生物相容性高的自由基反应条件，发展能够选择性识别和后修饰异戊烯基的新生物正交反应体系，实现对tRNA特定异戊烯基化位点和游离的i6A的动态可视化修饰，为研究发展具有实际应用价值的异戊烯基化RNA与其互作蛋白的作用模型提供重要科学基础，从而为发现i6A RNA新的生物学功能提供关键线索。

In this project, we will focus on the highly efficient, selective and sensitive chemical regulation and labeling of i6A, a chemical modification with important biological functions in tRNA. This method breaks through the traditional research methods through gene knockout/protein overexpression, but will develop selective oxidative demodification transformations based on the high activity of isoprenyl group at N6 position. By controlling the intracellular content of i6A will facilitate the study between the expression level and the cell phenotype. We will also screen biocompatible free radical reactions as new bioorthogonal transformations for studying the precise position of i6A tRNA and free i6A. This approach will also establish effective fishing technologies for the selective recognition of binding proteins pertaining to i6A, which will provide several key clues for discovering the new biological functions of i6A RNA.