Th17细胞中RORγt基因调控序列的鉴定

Th17细胞在感染和肿瘤等疾病中发生着非常重要的作用，RORγt为目前已确定的对Thl7细胞分化起决定性作用的特异性转录因子，因此对于RORγt基因的深入研究对于明确人体多种疾病发病机制及防治措施意义重大。目前对于RORγt基因下游功能研究比较多，但有关RORγt基因本身表达的调控研究还只是鉴定了该基因的启动子序列，对于调节启动子活性的更高级调控元件如增强子和沉默子的研究却未见报道，而后者对于基因在不同状态的细胞中的表达至关重要。因此本研究拟利用表观遗传学方法对Th17细胞基因组进行ChIP-Seq 扫描得到候选调节性序列，再利用pGL3-Luc报告系统、BAC-Luc报告系统对RORγt基因候选调节片断进行功能鉴定，筛选出真正调节性基因或片断。本研究有望找到Th17细胞形成的决定因素，为其作用机制的解码提供重要数据；且本研究结果本身也可能成为各种涉及到Th17细胞的疾病的潜在干预靶标。

Th17 cells play a key role in tumorogenesis, transplantation, autoimmune diseases and viral infection. RORγt gene has been proven to be the sole phenotypic and functional marker of Th17 to date. Thus, the study on RORγt gene expression will provide the key data on clarification of the mechanism of many related diseases and the establishment of strategies to control these diseases. Up to date, there have been many reports on the function of RORγt protein on its downstream molecules. However, there is rare research on the regulation of the expression of RORγt gene itself, and the long range regulation(such as enhancers and silencers) of this gene expression is almost untouched by now. Although no relative report at present, the long range regulation of RORγt gene expression should be of great importance and the outcome of many gene expression is determined dramatically by this kind of regulation. Hence, in this study, we will focus on this field with epigenetic strategy. We will screen the candidate regulatory elements by ChIP-Sequencing methods, then validate the real function and spatial correlation of these candidates by Luc reporter system . The expected results from this study will explore the determinant factors about Th17 formation and provide important data to dissect the mechanism underlying Th17 function. Furthermore, the results themselves could be the targets to control many Th17-related diseases.