Inactivation of the murine N-ras gene by gene targeting.
The mammalian ras genes have been implicated in the genesis of a wide variety of tumors. Although it is likely that they play an essential role in signal transduction, their specific function is still unknown. To initiate a genetic analysis of the ras genes in mammals we inactivated the N-ras gene in murine embryonic stem cells by gene targeting. The frequency of integration at the N-ras locus of our targeting vector being low (of the order of 1/5000 transfectants), we used a positive/negative selection followed by an analysis of individual colonies in order to minimize the in vitro manipulation of the embryonic stem cells. Using this approach, we isolated two clones of ES cells with one inactivated N-ras allele. These cells have no distinctive phenotype either in vitro or in vivo in chimeric mice.
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Laboratoire d'Oncologie Moléculaire, CNRS UA 1158, Institut Gustave Roussy, Villejuif, France.
Dautry
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D000483:Alleles; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002678:Chimera; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation; D011905:Genes, ras; D005822:Genetic Vectors; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D013234:Stem Cells / Q000502:physiology
D009838:Oligodeoxyribonucleotides; D004247:DNA