RNA解旋酶D1PAS1在雄性减数分裂中的作用机制研究

RNA解旋酶DDX3家族基因变异可能导致人类非梗阻性无精症，并与多种器官发育缺陷有关。然而，DDX3家族蛋白在哺乳动物雄性配子发生中的作用机制尚缺乏系统深入的研究，这有碍其人类致病位点的筛查和论证。有趣的是，Ddx3y敲除和Ddx3x/Ddx3y双敲的小鼠生精细胞发育正常。据此推断，有个同家族常染色体上的逆转座基因D1pas1可能在小鼠中发挥了类似功能。本项目通过构建点突变和敲除两种小鼠模型，利用CLIP-seq、ChIP-seq、体外酶活等多种实验方法和生物信息学分析手段，努力阐明D1PAS1在雄性减数分裂中的具体作用机理，力求揭示D1PAS1协同调控转录和转录后基因表达的独特方式。在此基础上，对无精症外显子测序得到的突变在蛋白活性方面进行初步的致病性探讨。本研究将立足基础研究，着眼临床意义，期待为探寻男性不育的遗传病理提供新的科学依据或理论。

Pathogenic variants in the DDX3 (RNA helicase) gene family lead to nonobstructive azoospermia in human. These variants are also responsible for multiple organ development defects. However, the mechanism of DDX3 family proteins in mammalian male gametogenesis is still lack of systematic and in-depth study, which hinders the screening and demonstration of clinical pathogenic variations. Interestingly, the mouse spermatogenic cells with Ddx3y knockout or Ddx3x / Ddx3y double knockout were normal. Thus we believe that D1pas1, which is the autosomal paralog of Ddx3x, may play a similar role in mice. By establishing point mutation and global knockout mouse models, using CLIP-seq，ChIP-seq，biochemical assay and bioinformational analysis, the project tries to clarify the role of D1PAS1 in male meiosis, and tries to reveal the unique way of D1PAS1 coordinating transcription and post-transcriptional regulation. On this basis, we will study the pathogenicity of the mutation obtained from whole exome sequence data from azoospermia patients. Based on basic research and clinical significance, this study is expected to provide a new scientific evidence or theory for exploring the genetic pathology of male infertility.