REGULATION OF ENDOTHELIN-1 GENE EXPRESSION

内皮素-1 基因表达的调节

• 批准号: 3366171
• 负责人: THOMAS QUERTERMOUS
• 金额: $ 3.91万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3366171
• 关键词: cardiovascular disorder; gene expression; genetic models; genetic promoter element; genetic transcription; genetically modified animals; growth factor; laboratory mouse; peptides; reporter genes; vascular endothelium; vasoconstrictors; vasomotion

Endothelin-1 (ET-1) is a 21-amino acid peptide synthesized in endothelial cells. It is both a potent vasoconstrictor and growth factor. The role of ET-1 in vascular physiology and its involvement in vascular disease is unknown. Aims of the proposed work include the use of genetic studies to better understand the function of ET-1, the use of this gene as a model system for the study of endothelial cell transcription, and the use of ET-1 regulatory sequences for endothelial cell targeting in transgenic animals. We will elucidate the cis-acting DNA elements of the ET-1 gene in vitro by transfection of ET-1 sequences linked to a reporter gene. These experiments will be complemented by analogous studies in vivo using transgenic mice which carry ET-1-lacZ transgenes. In vitro footprinting and gelshift analysis will allow characterization of trans-acting proteins which bind these sequences. ET-1 cis-acting sequences which appear to be specific for endothelial cell transcription will be employed in the purification and cloning of DNA binding proteins. Elements of the ET-1 promoter which provide high level endothelial cell expression in transgenic animals will be utilized to develop lines of transgenic mice which express either increased or attenuated levels of ET-1 in the blood vessel wall. These mice will be evaluated for alterations in blood pressure and predisposition for vascular disease. The availability of a clone for an endothelial cell-specific trans-acting factor will make possible studies investigating the development and phenotypic diversity of this important cell type. The availability of constructs which direct endothelial cell expression in transgenic mice will allow an approach to the generation of mouse models of vascular disease.

内皮素-1(ET-1)是在体内合成的一种由21个氨基酸组成的多肽。 内皮细胞。它既是一种有效的血管收缩药，也是一种生长 因素。ET-1在血管生理学中的作用及其在血管内皮细胞损伤中的作用 血管疾病是未知的。拟议工作的目标包括使用 为了更好地了解ET-1的功能，使用 该基因作为研究内皮细胞的模型系统 转录，以及ET-1调节序列在内皮细胞中的应用 转基因动物的细胞靶向。 我们将阐明ET-1基因的顺式作用DNA元件在 体外将ET-1序列与报告基因相连。这些 实验将得到体内类似研究的补充，使用 携带ET-1-lacZ转基因的转基因小鼠。体外足迹研究 而凝胶位移分析将允许表征反式作用 结合这些序列的蛋白质。ET-1顺式作用序列 似乎是内皮细胞特异的转录将被使用 在DNA结合蛋白的纯化和克隆中的应用。中的元素 ET-1启动子在血管内皮细胞高水平表达 转基因动物将被用于培育转基因小鼠品系 它们表达血液中ET-1水平的升高或减弱 血管壁。这些小鼠将接受血液变化的评估 血管疾病的压力和易感性。 内皮细胞特异性克隆的可用性 反式作用因子将使研究成为可能 这一重要细胞类型的发育和表型多样性。这个 指导血管内皮细胞表达的构建物的可用性 转基因小鼠将为建立小鼠模型提供一种途径 血管疾病。