The protein U5-52K is required for T cell homeostasis by maintaining splicing efficiency

U5-52K is a nuclear protein present in the U5 snRNP spliceosome subunits and it is essential for the mouse embryo development. In fact its absence blocks embryo at the developmental stage at E 10.5 with subsequence reasbsorbtion. Therefore, we used a CD4-Cre-knockout strategy to abrogate U5-52K in T cells, allowing us to delineate the consequences of splicing machinery interferences with T cell homeostasis. Several Differentially Spliced Variants and Differentially Expressed Genes were detected in T cells lacking U5-52K indicating its involvement in gene transcription regulation. Overall design: The RNA extracted from CD4+ isolated T cells from spleen and lymph nodes from three cKO mice (U5-52Kf/f CD4-Cretg) and three control mice (Control: U5-52Kwt/f CD4-Cretg and U5-52Kf/f CD4-Cre-) were sequenced using RNA-seq technology.

U5-52K是一种定位于U5小核核糖核蛋白（U5 snRNP）剪接体亚基的核蛋白，对小鼠胚胎发育至关重要。实际上，该蛋白的缺失会使胚胎阻滞于E10.5发育阶段，随后发生胚胎重吸收。为此，我们采用CD4-Cre敲除（CD4-Cre-knockout）策略敲除T细胞中的U5-52K，以此解析剪接机器功能异常对T细胞稳态的影响。在缺失U5-52K的T细胞中，我们检测到多种差异剪接变体（Differentially Spliced Variants）与差异表达基因（Differentially Expressed Genes），提示其参与基因转录调控过程。整体实验设计：从3只条件性敲除（conditional knockout, cKO）小鼠（基因型为U5-52Kf/f CD4-Cretg）及3只对照小鼠（对照组基因型为U5-52Kwt/f CD4-Cretg 与 U5-52Kf/f CD4-Cre-）的脾脏和淋巴结中分离CD4+ T细胞，提取RNA后采用RNA测序（RNA-seq）技术进行测序。