THE FUNCTION OF RE-1/REST IN CRH REGULATION

探讨核因子限制性元件沉默转录因子(REST)通过位于CRH基因第一内含子的限制性元件1(RE-1元件)负调控CRH基因表达的可能性。A)通过凝胶迁移率改变分析(EMSA)证实REST与CRH RE-1序列的特异性结合，为REST介导的CRH转录调控奠定了基础。2)体外转录实验表明，RE-1可通过REST介导CRH的转录抑制。II.通过小鼠敲入缺少RE-1的CRH基因的一个版本，验证RE-1元件在体内介导CRH基因重要转录调控的假设。

To establish the potential for the nuclear factor, Restrictive Element Silencing Transcription factor (REST), to negatively regulate CRH gene expression via the Restrictive Element 1 (RE-1 element) located in the first intron of the CRH gene. A) Demonstrate specific REST binding to the CRH RE-1 sequence in vitro, by Electrophoretic Mobility Shift Assay (EMSA) to establish the possibility for RE-1 mediated CRH transcriptional regulation by REST. B) Show by in vitro transcriptional assays that RE-1 can mediate CRH transcriptional repression by REST. II. To test the hypothesis that the RE-1 element mediates important in vivo transcriptional regulation of the CRH gene by mouse knock-in of a version of the gene lacking the RE-1.