GENETIC ANALYSIS OF FGF-5 PROTO-ONCOGENE FUNCTION

FGF-5原癌基因功能的遗传分析

• 批准号: 3328780
• 负责人: MITCHELL GOLDFARB
• 金额: $ 9.08万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3328780
• 关键词: RNA splicing; alleles; cell differentiation; developmental genetics; fibroblast growth factor; gene expression; gene mutation; genetic manipulation; genetic recombination; genetic translation; genetically modified animals; in situ hybridization; inbreeding; laboratory mouse; microinjections; neoplasm /cancer genetics; protooncogene; tissue /cell culture; transfection; transposon /insertion element

The FGF-5 proto-oncogene specifies a secreted growth factor which bears sequence homology to the prototypic fibroblast growth factors. FGF-5. is expressed in several embryonic settings, in mature brain, and in some neoplastic cells. This application proposes a series of genetic experiments to determine many of FGF-5's distinct functions. We shall disrupt one or both alleles of the FGF-5 gene in murine totipotent embryonic stem (ES) cells by homologous recombination. FGF-5 negative ES cells will be analyzed for differentiation potential as embryoid bodies in vitro and as tumors in vivo. FGF-5 hemizygous ES cells will be injected into blastocysts to derive chimeric mice and establish a disrupted allele in the germ line. Inbreeding will be performed to determine the developmental abnormalities in FGF-5 negative embryos. We shall also establish transgenic mice bearing human FGF-5 genes which are expressed in a subset of the developmentally characteristic sites or which bear structural mutations in a domain of currently unknown function. Mating these transgenic strains with FGF-5 hemizygous mice and backcrossing progeny with the hemizygous strain will generate embryos carrying the transgenes as sole functional allele. The aberrant phenotypes of these embryos will further dissect FGF-5 function. Lastly, we shall generate mice carrying human FGF-5 transgenes mutated at sites known to increase the translation efficiency of FGF-5 MRNA. These mice should overexpress FGF-5 protein without ectopic expression, and resulting abnormalities will also help determine FGF-5 function. These mice shall also be monitored for cancer predisposition.

FGF-5原癌基因指定一种分泌生长因子