GENETIC ANALYSIS OF FGF-5 PROTO-ONCOGENE FUNCTION
GENETIC ANALYSIS OF FGF-5 PROTO-ONCOGENE FUNCTION
批准号:
3328778
负责人:
MITCHELL GOLDFARB
金额:
$22.51万
依托单位国家:
美国
项目类别:
财政年份:
1990
资助国家:
美国
项目状态:
已结题
起止时间:
1990-08-01 至 1993-07-31
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
中文摘要
FGF-5原癌基因指定一种分泌的生长因子,
与原型成纤维细胞生长因子序列同源。FGF-5。是
在几种胚胎环境中表达,在成熟的大脑中表达,在一些
肿瘤细胞这个应用程序提出了一系列的遗传实验
以确定FGF-5的许多独特功能。我们将破坏一个或
小鼠全能胚胎干细胞(ES)中FGF-5基因的两个等位基因
同源重组的细胞。将分析FGF-5阴性ES细胞
对于体外作为胚状体的分化潜力和作为肿瘤的分化潜力,
vivo. FGF-5半合子ES细胞将被注射到胚泡中,以衍生
嵌合小鼠,并在生殖系中建立破坏的等位基因。近亲繁殖
以确定FGF-5的发育异常
阴性胚胎我们还将建立转基因小鼠,
FGF-5基因在发育过程中的一个亚群中表达,
特征性位点或在结构域中具有结构突变的
目前未知的功能将这些转基因菌株与FGF-5交配
半合子小鼠和具有半合子品系的回交后代将
产生携带转基因作为唯一功能等位基因的胚胎。的
这些胚胎的异常表型将进一步剖析FGF-5的功能。
最后,我们将产生携带人FGF-5转基因的小鼠,
已知增加FGF-5 mRNA翻译效率的位点。这些
小鼠应过表达FGF-5蛋白而无异位表达,
由此产生的异常也将有助于确定FGF-5的功能。这些小鼠
还应监测癌症易感性。
英文摘要
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.
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