FTO通过m6A去甲基化修饰的GADD45B调控山羊成肌细胞分化的机制研究

我国地方品种山羊普遍存在着产肉性能欠佳的问题，阐明肌肉发育机理至关重要，其中m6A RNA甲基化修饰在调控肌肉生长发育中发挥重要作用。申请人前期通过对山羊成肌细胞m6A-seq和RNA-seq的联合分析，筛选到关键的m6A去甲基化酶FTO及其候选靶基因GADD45B，其可能与山羊成肌细胞分化存在相关性。因此，本项目拟以山羊成肌细胞为对象，首先通过MeRIP-qPCR、挽救试验等，分析FTO是否通过m6A去甲基化修饰靶向调控GADD45B的表达；其次，探究FTO是否通过调控GADD45B表达进而影响山羊成肌细胞分化；最后，通过干扰/过表达FTO和GADD45B、抑制或激活候选通路p38 MAPK-mTOR，阐明FTO通过m6A修饰的GADD45B调控山羊成肌细胞分化的作用机制。本项目瞄准产业问题，探索山羊肌肉发育的调控机制，对推动肉用山羊育种具有重要的科学意义。

Local goat breeds in China generally have the problem of poor meat production performance. It is important to clarify the mechanism of muscle development. Among of them, m6A RNA methylation modification plays an important role in the regulation of muscle growth and development. Through a comprehensive analysis of our previous m6A-seq and RNA-seq of goat myoblast, the applicant has screened the m6A demethylase FTO and its candidate target gene GADD45B, and they may be related to the myoblast differentiation of goat. Therefore, this project uses goat myoblasts as the experimental subjects. First, the targeting relationship between m6A demethylase FTO and GADD45B is explored through MeRIP-qPCR and rescue test. Secondly, the biological function of FTO in mediating the expression of GADD45B in myoblast differentiation is analyzed. Finally, the regulatory mechanism of FTO-mediated GADD45B m6A modification on goat myoblast differentiation is verified from both positive and negative aspects, by interference or overexpression of FTO or GADD45B, and inhibit or activate p38 MAPK-mTOR pathway. This project aims at the industry problem and explores the regulation mechanism of goat muscle development, which has important practical significance for accelerating goats breeding.