Functional Analysis of Novel Genes in Eye Development and Vision

1: AMD与补体因子H （CFH）。CFH的常见序列变异在决定老年性黄斑变性（AMD）的易感性中起主要作用。利用酵母2-杂化文库和CFH的amd相关结构域7的诱饵构建物，我们已经确定了CFH在人RPE/脉络膜中结合的潜在重要靶点。这些结果已通过免疫荧光定位和天然和重组蛋白结合实验得到证实。质谱正在被用来鉴定与AMD中蛋白质沉积形成有关的复合物的进一步成分。这为药物设计干预AMD的进展提供了潜力。AMD中应力相关过程的细胞培养模型也正在研究中。

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.