TRANSGENIC ANAYLSIS OF RETINAL DEVELOPMENT IN VERTEBRATE

脊椎动物视网膜发育的转基因分析

• 批准号: 2885608
• 负责人: James M Fadool
• 金额: $ 25.8万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2004-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2885608
• 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

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 陷阱技术将得到完善，随后将被系统地利用， 定义了数百个在视网膜发育中重要的基因， 斑马鱼该技术是一个强大的适应良好的特点， 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的 用无毒荧光染料标记的转座子的表达，即，绿色 荧光蛋白（GFP）。转座子的相对有效整合 已经得到证明，超过5%的集成事件是 预计将在内源性增强子的控制下导致GFP表达 选择细胞和组织的群体。在其中GFP是 将鉴定在视网膜或其它组织中选择性表达的蛋白， 进行更详细的发育、形态和遗传分析， 首席研究员和其他人。在果蝇中， 预期转座事件的发生会干扰正常的基因表达， 导致突变或数量变异。增强子陷阱的这个特征 是特别重要的，因为它应该能够对 视网膜的细微异常，否则会逃脱检测。