THE FUNCTION OF RE-1/REST IN CRH REGULATION

RE-1/REST 在 CRH 调节中的作用

• 批准号: 6139369
• 负责人: KIM A SETH
• 金额: $ 2.04万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-16 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6139369
• 关键词: corticotropin releasing factor; gel mobility shift assay; gene targeting; genetic regulatory element; hormone regulation /control mechanism; laboratory mouse; transcription factor

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.

为了确定核因子限制性元件沉默转录因子（REST）通过位于CRH基因第一个内含子的限制性元件1 （RE-1元件）负性调节CRH基因表达的潜力。A)通过EMSA （electrophotic Mobility Shift Assay）在体外证明REST与CRH RE-1序列的特异性结合，以确定REST对RE-1介导的CRH转录调节的可能性。B)通过体外转录实验表明RE-1可以通过REST介导CRH转录抑制。2。通过敲入缺乏RE-1的基因版本，验证RE-1元件介导CRH基因在体内重要转录调控的假设。