Genetic Screen for Retinal Degeneration Mutants in Drosophila

描述(申请人提供)：利用果蝇黑腹果蝇的遗传学方法，我们发现编码果蝇肌动蛋白结合蛋白cofilin/ADF同源物的基因Twinstar(TSR)在眼睛中具有生理作用，因为TSR具有光依赖型视网膜变性(RD)突变表型。我们认为TSR代表了一大组生存所需的基因，这些基因具有未知的RD突变表型。为了测试这一点，对果蝇中的RD突变体进行了RNAi筛选。初步数据显示，这种方法将是成功的。这项建议的目标是筛选8,000个RNAi诱导系以寻找RD突变表型。一旦确定，新的RD突变体将被鉴定以确定视网膜最早的可见缺陷，RD是否依赖于光，以及变性是凋亡性还是坏死性。果蝇和人类的RD基因有显著重叠，表明它们具有共同的功能。识别更多的果蝇RD突变体将把人类同源物标记为与视网膜疾病相关的候选。许多RD基因也有可能在果蝇的眼睛中具有有趣的生物学功能。识别这些新的RD基因将把新的研究人员带入RD领域，从而更好地了解视力和视网膜疾病。 与公共健康相关：最近对果蝇的研究表明，导致果蝇视网膜退化的许多基因的突变也会导致人类的视网膜退化，这表明这是一种常见的功能。这项资助提出了一种基因筛查，以确定果蝇中具有视网膜变性突变表型的其他基因。确定这些基因的作用和分子功能将有助于加深对视力和视网膜疾病的理解。

DESCRIPTION (provided by applicant): Using a genetic approach with the fruit fly Drosophila melanogaster, we have found that the gene twinstar (tsr), which encodes the Drosophila homologue of the actin-binding protein cofilin/ADF, has a physiological role in the eye, as tsr has a light-dependent retinal degeneration (RD) mutant phenotype. We believe tsr represents a large group of genes that are required for viability that have an unidentified RD mutant phenotype. To test this, an RNAi screen for RD mutants in the fly was started. Preliminary data shows that this approach will be successful. The goal of this proposal is to screen 8,000 RNAi inducible lines for an RD mutant phenotype. Once identified, the new RD mutants will be characterized to determine the earliest visible defect in the retina, if the RD is light dependent, and if the degeneration is apoptotic or necrotic. There is significant overlap between RD genes in the fly and humans, indicating common function. Identifying additional Drosophila RD mutants will mark the human homologues as candidates for being associated with retinal diseases. It is also likely that many of the RD genes will have interesting biological functions in the Drosophila eye. Identifying these new RD genes will bring new researchers into the field of RD, leading to a better understanding of vision and retinal disease. PUBLIC HEALTH RELEVANCE: Recent studies in Drosophila have shown that mutants of many genes that cause retinal degeneration in the fly also cause retinal degeneration in humans, suggesting a common function. This grant proposes a genetic screen to identify additional genes in Drosophila that have a retinal degeneration mutant phenotype. Characterizing the roles and molecular functions of these genes will add to an understanding of vision and retinal disease.