An endothelial-specific switch in gene expression.

基因表达中的内皮特异性开关。

• 批准号: 6835529
• 负责人: Jill M Johnsen
• 金额: $ 5.44万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835529
• 关键词: DNA footprinting; animal genetic material tag; artificial chromosomes; galactosyltransferases; gel mobility shift assay; gene expression; genetic regulation; genetic regulatory element; genetically modified animals; intermolecular interaction; laboratory mouse; nucleic acid sequence; nucleic acid structure; postdoctoral investigator; protein localization; vascular endothelium; von Willebrand' s disease

DESCRIPTION (provided by applicant): The inbred mouse strain RIIIS/J is a model for type 1 von Willebrand Disease (VWD), the most common bleeding disorder in humans. We previously found that the cause of low VWF in RIIIS/J is a regulatory mutation, Mvwfl, in the gene encoding an N-acetylgalactosaminyltransferase, GALGT2. Mvwfl causes a tissue-specific switch from the intestinal epithelial expression pattern observed in most mouse strains to a vascular endothelial pattern, resulting in aberrant post-translational modification of VWF and accelerated clearance. The precise cis-regulatory elements responsible for this remarkable switch have not yet been identified. We hypothesize that characterization of the regulatory element(s) responsible for the RIIIS/J switch will significantly advance our understanding of vascular and intestine specific gene expression programs. This project will utilize direct sequence analysis and engineered bacterial artificial chromosome systems (BACs) to identify the genomic regions responsible for the switch. Comparative sequence analysis of different Mvwfl mouse strains should determine the minimum length of the shared Mvwfl haplotype block. Chimeric RIIIS/J:C57BL6/J BACs will be engineered by swapping fragments of the RIIIS/J candidate sequence into a C57BL/6J BAC that is known to exhibit intestinal epithelial Galgt2 expression. Transgenic mice will be generated and tested for tissue patterns of Galgt2 expression. Constructs that confer the RIIIS/J Galgt2 expression pattern should define the region containing the critical regulatory element(s), which will be further characterized by additional BAC mutations. Our findings should provide new insight into the cis-regulatory elements required for endothelial gene expression, with potential broad implications for other tissue specific gene expression programs and vascular gene therapy.

描述（由申请人提供）：近交小鼠品系 RIIIS/J 是 1 型冯维勒布兰德病 (VWD) 的模型，这是人类最常见的出血性疾病。我们之前发现 RIIIS/J 中低 VWF 的原因是编码 N-乙酰半乳糖氨基转移酶 GALGT2 的基因中的调节突变 Mvwfl。 Mvwfl 导致组织特异性从大多数小鼠品系中观察到的肠上皮表达模式转变为血管内皮模式，导致 VWF 异常翻译后修饰并加速清除。造成这种显着转变的精确顺式调控元件尚未确定。我们假设负责 RIIIS/J 开关的调控元件的表征将显着增进我们对血管和肠道特异性基因表达程序的理解。该项目将利用直接序列分析和工程细菌人工染色体系统（BAC）来识别负责转换的基因组区域。不同 Mvwfl 小鼠品系的比较序列分析应确定共享 Mvwfl 单倍型块的最小长度。将通过将 RIIIS/J 候选序列的片段交换到已知表现出肠上皮 Galgt2 表达的 C57BL/6J BAC 中来工程化嵌合 RIIIS/J:C57BL6/J BAC。将生成转基因小鼠并测试 Galgt2 表达的组织模式。赋予 RIIIS/J Galgt2 表达模式的构建体应定义包含关键调控元件的区域，该区域将通过额外的 BAC 突变进一步表征。我们的研究结果应该为内皮基因表达所需的顺式调控元件提供新的见解，对其他组织特异性基因表达程序和血管基因治疗具有潜在的广泛影响。