CIS-REGULATORY MECHANISMS UNDERLYING RETINOPATHY

视网膜病变的 CIS 调节机制

• 批准号: 9288077
• 负责人: SHIMING CHEN
• 金额: $ 34.31万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9288077
• 关键词: Affect; Age of Onset; Binding; Binding Sites; Biological Assay; ChIP-seq; Chromatin; Computer Simulation; DNA; DNA-Protein Interaction; Data; Development; Disease; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genome; Genomic Library; Genomic medicine; Genomics; Global Change; Goals; Human; Knock-in; Knock-in Mouse; Lead; Leber' s amaurosis; Link; Maintenance; Measurement; Measures; Methods; Modeling; Molecular; Mus; Mutation; Pathogenesis; Pathologic; Pathology; Pattern; Phenotype; Photoreceptors; Proteins; Reporter; Reporter Genes; Retina; Retinal Diseases; Retinitis Pigmentosa; Role; Severities; Site; Technology; Testing; Training; base; design; disease-causing mutation; genome-wide; in vivo; mouse model; mutant; predictive modeling; protein protein interaction; targeted treatment; therapy development; transcription factor

Project Summary    Different human mutations in the photoreceptor-specific gene Crx are linked with multiple retinopathies that vary in their severity and age of onset. How different mutations in this single gene lead to different pathologies is not well understood. Because CRX is a transcription factor, these disease mutations must act by altering the ability of CRX to regulate gene expression. Our goal is to understand how different classes of CRX mutations modify its regulatory function. To achieve this, we will apply a recently developed massively parallel reporter gene technology to systematically and comprehensively measure CRX regulatory function in live retina from multiple mouse knock-in models that carry human CRX disease mutations. We will use the data to train and test a mechanistic, quantitative model that describes how CRX mutations alter protein-DNA and protein-protein interactions to modify gene regulation. In a complementary approach, we will use the knock-in mouse models to determine the effects of CRX disease mutations on its in vivo genome-wide binding, and on the binding of the cooperatively interacting transcription factors OTX2, NRL, and NR2E3. Using these two approaches, we aim to understand how different classes of CRX disease mutations modify gene regulation in photoreceptors, and discover new mechanisms of disease pathogenesis. Our results will provide a strong basis for classifying new CRX mutations, and for designing targeted therapies to treat different genetic forms of retinopathy.

项目摘要   光感受器特异性基因Crx中的不同人类突变与 严重程度和发病年龄不同的多种视网膜病变。多么不同 在这个单一基因中的突变导致不同的病理还没有很好地理解。 由于CRX是一种转录因子，这些疾病突变必须通过改变CRX基因的表达来起作用。 CRX调节基因表达的能力。我们的目标是了解 CRX突变改变了其调节功能。为了实现这一点，我们将应用 最近开发了大规模平行报告基因技术， 多只小鼠活体视网膜CRX调节功能的综合测定 携带人类CRX疾病突变的基因敲入模型。我们将用这些数据来训练 并测试一个描述CRX突变如何改变 蛋白质-DNA和蛋白质-蛋白质相互作用以改变基因调控。中 互补的方法，我们将使用敲入小鼠模型来确定 CRX疾病突变对其体内全基因组结合的影响， 协同相互作用的转录因子OTX 2、NRL和NR 2 E3的结合。 使用这两种方法，我们的目标是了解不同类别的CRX 疾病突变改变了光感受器的基因调控，并发现了新的 疾病的发病机制。我们的研究结果将为 分类新的CRX突变，并设计靶向治疗，以治疗不同的 视网膜病变的遗传形式。