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TRANSGENIC ANAYLSIS OF RETINAL DEVELOPMENT

视网膜发育的转基因分析

基本信息

批准号：
6663680
负责人：
James M Fadool
金额：
$ 28.87万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-30 至 2006-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6663680
关键词：
developmental genetics gene expression gene targeting genetic enhancer element genetic techniques genetically modified animals green fluorescent proteins phenotype retina transposon /insertion element vertebrate embryology zebrafish

项目摘要

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.

描述（改编自申请人的摘要）：高通量增强剂陷阱技术将被完善并随后被系统地利用定义了数百个对视网膜发育很重要的基因斑马鱼。该技术是对已充分表征的技术的强大改编 P-转座子方法在剖析遗传学方面非常有效果蝇发育。在这种情况下，该方法依赖于选择性用无毒荧光染料（即绿色）标记的转座子的表达荧光蛋白（GFP）。转座子的相对有效的整合已经被证明，超过 5% 的整合事件是预计在内源性增强子的控制下导致 GFP 表达选择细胞和组织群体。含有 GFP 的鱼系将鉴定在视网膜或其他组织中选择性表达的保存用于更详细的发育、形态测量和遗传分析首席研究员和其他人。与果蝇一样，很大一部分预计转座事件会扰乱正常基因表达并导致突变或数量变异。增强子陷阱的这一特点特别重要，因为它应该能够进行细粒度的分析视网膜上的细微异常可能会逃避检测。