Zbtb1基因在T细胞发育过程中调控IL-7Rα表达的功能和机制研究

BTB-ZF转录因子家族的成员Zbtb1对T细胞发育至关重要。在我们的前期研究工作中发现，Zbtb1在小鼠T细胞发育过程中能调控IL-7Rα的表达。IL-7信号由IL-7受体传导，它对T细胞成熟和分化至关重要。本项目针对Zbtb1基因突变导致T细胞发育缺陷的现象，从Zbtb1基因调控IL-7Rα的表达方面入手，拟采用在体外和体内水平过表达Zbtb1蛋白水平的方法，确定Zbtb1基因对IL-7Rα表达的抑制作用。通过Biotin ChIP-seq技术，寻找Zbtb1结合的与IL-7Rα表达相关的顺式作用元件。在小鼠DN胸腺细胞系(D1)中通过CRISPR-Cas9技术突变与Zbtb1结合的顺式作用元件的关键位点，在野生型和突变型D1细胞系中检测过表达Zbtb1对IL-7Rα表达调控的差异，可阐明Zbtb1调控IL-7Rα表达的分子机制。研究结果将完善Zbtb1基因调控T细胞发育的功能和机制。

Zbtb1 is a member of the zinc finger and BTB domain containing protein family of transcriptional factors, which is critical for T cell development. In previous study, we found that Zbtb1 can regulate the expression of IL-7Rα during T cell development of mouse. IL-7 signaling is transduced by the IL-7 receptor, which is essential for T cell maturation and differentiation. As developmental defects of T cells induced by Zbtb1 mutation, the present study aimed to investigate the regulation of IL-7Rα gene expression by Zbtb1 gene. We’re going to overexpress Zbtb1 gene in vitro and in vivo, which can confirm the inhibitory effect of Zbtb1 on IL-7Rαexpression. The binding of Zbtb1 to cis-element related to IL-7Rαexpression will be identified by Biotin ChIP-seq technique. The key sites of cis-element for Zbtb1 binding will be mutated by CRISPR-Cas9 technique in a mouse DN thymocyte cell line (D1). The differential regulation of IL-7Rαwill be tested in wild type and mutant D1 cell line by overexpression of Zbtb1, which can illustrate the molecular mechanism for IL-7Rαregulation by Zbtb1. The results for this study will shed light on the function and mechanism of Zbtb1 on T cell development.