CONSECUTIVE INACTIVATION OF BOTH ALLELES OF THE PIM-1 PROTOONCOGENE BY HOMOLOGOUS RECOMBINATION IN EMBRYONIC STEM-CELLS
CONSECUTIVE INACTIVATION OF BOTH ALLELES OF THE PIM-1 PROTOONCOGENE BY HOMOLOGOUS RECOMBINATION IN EMBRYONIC STEM-CELLS
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DOI：
10.1038/348649a0
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发表时间：
1990-12-13
期刊：
影响因子：
64.8
通讯作者：
BERNS, A
中科院分区：
文献类型：
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作者：
RIELE, HT;MAANDAG, ER;BERNS, A
在小鼠生殖系中，特定基因可能失活或突变。突变的表型结果可以为该基因在哺乳动物机体发育和维持中的功能提供关键信息。这个过程需要在胚胎干细胞中进行同源重组，在与受体囊胚融合后，产生嵌合小鼠，这些小鼠可以将突变基因传递给它们的后代。近亲繁殖可以产生携带两个等位基因突变的小鼠，从而可以对隐性突变进行表型分析。除了缺乏特定基因功能的小鼠外，携带正常表达基因的零等位基因的细胞系可以用于评估该基因的功能。这些细胞系既可以从纯合子动物中获得，也可以通过在培养细胞中通过同源重组使两个等位基因连续失活而产生，如果突变在胚胎发育早期是致命的。在这里，我们通过在胚胎干细胞中有效地连续灭活pim-1原癌基因的两个等位基因来说明后一种方法的可行性。
SPECIFIC genes can be inactivated or mutated in the mouse germ line1. The phenotypic consequences of the mutation can provide pivotal information on the function of the gene in development and maintenance of the mammalian organism. The procedure entails homologous recombination in embryonic stem cells, which, on fusion to recipient blastocysts, give rise to chimaeric mice that can transmit the mutant gene to their offspring. Inbreeding can then yield mice carrying the mutation in both alleles allowing the phenotypic analysis of recessive mutations. In addition to mice lacking a particular gene function, cell lines carrying null alleles of normally expressed genes can be instrumental in assessing the function of the gene. These cell lines can either be obtained from homozygous animals or, should the mutation be lethal early in embryonic development, be generated by consecutive inactivation of both alleles by homologous recombination in cultured cells. Here we illustrate the feasibility of this latter approach by the efficient consecutive inactivation of both alleles of thepim-1proto-oncogene in embryonic stem cells.