m6A甲基化修饰与哺乳动物卵巢和雌性生殖细胞发育密切相关，但机制不详。申请者前期通过构建模型小鼠，发现生殖细胞特异性Mettl14基因敲除小鼠不孕，且雌性生殖干细胞（FGSC）和各级卵泡数目减少。进一步研究显示Mettl14敲除导致FGSC的m6A甲基化水平下降，影响其组蛋白乙酰化修饰和细胞表型。本课题拟在此基础上，探索Mettl14通过组蛋白乙酰化修饰调控FGSC命运的机制。首先利用模型小鼠系统研究敲除Mettl14对小鼠卵巢和雌性生殖细胞发育的影响及机制，然后应用3D卵巢类器官、m6A-seq、RNC-seq等探究Mettl14对FGSC生物学特性影响和关键信号通路，最后通过靶向蛋白组质谱等手段研究Mettl14通过组蛋白乙酰化修饰影响FGSC命运决定的机制。项目的实施有助于深入理解m6A修饰在雌性生殖细胞发育中的关键作用，为卵子发生障碍等相关疾病诊治提供基础。

N6-methyladenosine (m6A) modification is closely related to the development of mammalian ovary and female germ cells, but the mechanism is not clear. Our previous studies showed that germ cell-specific knockout of Mettl14 (Mettl14 cKO) mice were infertile, and the number of female germline stem cells (FGSC) and follicles in the ovaries of the model mice decreased. Further studies demonstrated that Mettl14 knockout reduced the m6A level, and affected histone acetylation and cell phenotype of FGSC. On this basis, we will first systematically investigate the effect and mechanism of Mettl14 knockout on the development of mammalian ovary and female germ cells in mice useing Mettl14 cKO mice. And then study the impact of Mettl14 on the biological characteristics of FGSC and the key signaling pathway by 3D ovarian organoids , m6A-seq, RNC-seq, etc. Finally, we will explore the mechanism of Mettl14 regulates the fate of FGSC through histone acetylation by mass spectrometry–based targeted proteomics, etc. This project will shed light on understanding the key role of m6A modification in the development of female germ cells, and provide theoretical basis for the diagnosis and treatment of related diseases such as oogenesis disorders.