逆转座基因Nkapl在精子发生过程中的功能和机制研究

精子发生过程中特定发育阶段的转录因子和表观遗传蛋白协同调控基因的表达。NF-κB (Nuclear Factor Kappa B)家族的转录因子参与调节多种基因的转录，且存在激活和抑制两种模式。Nkapl (NF-κB-activating protein-like)是一个由X染色体逆转座至常染色体上的基因，特异地表达于雄性生殖细胞。我们通过CRISPR-Cas9技术构建了Nkapl基因敲除小鼠模型和NKAPL FLAG-HA标签鼠。Nkapl KO小鼠雄性不育，大部分生精管腔阻滞在圆形精子阶段。我们将深入分析Nkapl基因缺失小鼠生殖表型，探索其雄鼠不育的原因。RNA-seq、CoIP-质谱和染色质免疫沉淀联合高通量测序（ChIP-seq）联合分析将揭示NKAPL蛋白在基因组上的结合位点以及其调控基因表达的作用机理。

Stage-specific transcription factors coordinate with epigenetic factors to regulate gene expression in spermatogenesis. The NF-κB (Nuclear Factor Kappa B) family of transcription factors regulates transcription of a wide variety of genes through gene activation or repression. Nkapl (NF-κB-activating protein-like) is a gene that retrotransposes from X chromosome to autosomes, specifically expresses in male germ cells. We generated Nkapl KO and FLAG-HA tagged NKAPL mice by CRISPR-Cas9 technology. Nkapl KO male were infertile, with the majority of seminiferous tubules arrested at the round spermatid stage. In this study, we will investigate the phenotype of Nkapl KO males, and explore the related mechanism. Combined analyses including RNA-seq, CoIP-mass spectrometry and chromatin immunoprecipitation followed by sequencing (ChIP-seq) will uncover the genomic binding sites of NKAPL and address the mechanism through which NKAPL regulates gene expression.