TRANSGENIC ANAYLSIS OF RETINAL DEVELOPMENT

描述(改编自申请人的摘要)：一种高通量增强剂 圈闭技术将得到改进，随后将被系统地开发 定义数百个在视网膜发育中起重要作用的基因 斑马鱼。这项技术是对具有良好特征的 P-转座子方法在解剖人类基因组的遗传学方面非常有效 果蝇的发育。在这种情况下，该方法依赖于选择性的 无毒的绿色荧光标记转座子的表达 荧光蛋白(GFP)。转座子的相对有效整合 已经进行了演示，超过5%的集成活动是 有望在内源增强子控制下导致绿色荧光蛋白的表达 选择细胞和组织的群体。绿色荧光蛋白所在的鱼线 在视网膜或其他组织中选择性表达的基因将被鉴定并 保存以进行更详细的发育、形态测量和遗传分析 首席调查员和其他人。就像在果蝇中，相当一部分 转位事件的发生预计会扰乱正常的基因表达并 导致突变或数量变异。增强器陷阱的这一功能 尤其重要，因为它应该能够对 视网膜中的细微异常，否则就不会被发现。

DESCRIPTION (Adapted from the applicant's abstract): A high-throughput enhancer trap technique will be refined and subsequently exploited to systematically define hundreds of genes that are important in the retinal development of zebrafish. The technique is a powerful adaptation of well-characterized P-transposon methods that have been so effective in dissecting the genetics of Drosophila development. In this case, the method relies on the selective expression of transposons tagged with a nontoxic fluorochrome, i.e., green fluorescent protein (GFP). Relatively efficient integration of the transposon has been demonstrated, and more than 5 percent of integration events are expected to lead to GFP expression under the control of endogenous enhancers in select populations of cells and tissues. Lines of fish in which GFP is expressed selectively in retina or other tissues will be identified and preserved for more detailed developmental, morphometric and genetic analyses by the principal investigator and others. As in Drosophila, a significant fraction of transposition events is expected to disturb normal gene expression and to lead to mutations or quantitative variants. This feature of the enhancer trap is particularly significant because it should enable a fine-grained analysis of subtle abnormalities in retina that would otherwise escape detection.