Functional Analysis of Novel Genes in Eye Development and Vision

眼睛发育和视力新基因的功能分析

• 批准号: 8339778
• 负责人: graeme j wistow
• 金额: $ 63.26万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8339778
• 关键词: Age related macular degeneration; Aging; Animal Model; Binding; Bioinformatics; Carotenoids; Cataract; Cell Culture Techniques; Characteristics; Choroid; Complement Factor H; Complex; Deposition; Drug Design; Ear; Evaluation; Eye; Eye Development; Genes; Genomics; Growth; Hearing; Human; Hybrids; Immunofluorescence Immunologic; Knockout Mice; Libraries; Mass Spectrum Analysis; MicroRNAs; Modeling; Mus; Photoreceptors; Predisposition; Process; Protein Binding; Proteins; Recombinant Proteins; Retina; Retinal Cone; Retinoids; Role; Stress; Techniques; Tissues; Transgenic Mice; Variant; Vision; Yeasts; age related; aged; lens; mouse model; novel; promoter; research study; retinal rods

1: AMD and complement factor H (CFH). Common sequence variants of CFH have major roles in determining susceptibility to age-related macular degeneration (AMD). Using yeast 2-hybrid libraries for aged human RPE/choroid and retina and bait constructs for the AMD-related domain 7 of CFH we have identified potentially important targets for CFH binding in human RPE/choroid. These results have been confirmed by immunofluorescence localization and by using native and recombinant protein binding experiments. Mass spectroscopy is being used to identify further components of complexes that are implicated in formation of protein depositions in AMD. This has potential for drug-design to intervene in the progression of AMD. A cell-culture model for stress-related processes in AMD is also being investigated. 2: Retbindin is a novel protein of the retina. We have shown that retbindin is essentially specific for photoreceptors and that the human gene has separate cone and rod cell promoters. A transgenic mouse model shows suggests that retbindin has a role in control of retinoid and carotenoid levels in the retina. A knockout mouse has been created and is nearly ready for evaluation 3: Novel lens-specific genes with characteristics suggesting possible roles in age-related cataract have been identified and two mouse models with age-related cortical cataract have been created. 4: A miRNA cluster with high expression in eye and ear has been deleted in mouse, producing anew model of hearing and vision deficit.

1：AMD和补体因子H（CFH）。 CFH的常见序列变异在确定与年龄相关的黄斑变性（AMD）的敏感性方面具有重要作用。使用酵母2-杂交库来为CFH的AMD相关结构域7进行老化的人类RPE/脉络膜和视网膜以及诱饵构建体，我们已经确定了人类RPE/脉络膜中CFH结合的潜在重要目标。通过免疫荧光定位以及使用天然和重组蛋白结合实验证实了这些结果。质谱法被用来识别与AMD中蛋白质沉积形成有关的复合物的进一步成分。这具有药物设计的潜力，可以干预AMD的进展。还正在研究用于AMD中应力相关过程的细胞培养模型。 2：视网膜是视网膜的新型蛋白质。我们已经表明，乙蛋白基本上是对感光体的特异性特异性，并且人类基因具有单独的锥体和杆细胞启动子。转基因小鼠模型表明，视网膜蛋白在视网膜类维生素类和类胡萝卜素水平的控制中起作用。已经创建了淘汰鼠标，几乎可以评估 3：具有特征的新型镜头特异性基因，表明已经鉴定出可能在年龄相关性白内障中作用，并且已经创建了两种具有与年龄相关的皮质性白内障的小鼠模型。 4：在小鼠中删除了具有高度表达的miRNA簇，产生了听力和视力不足模型。