Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells.
Specific genes can be inactivated or mutated in the mouse germ line. 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 the pim-1 proto-oncogene in embryonic stem cells.
te Riele
H
H
Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam.
Maandag
E R
ER
Clarke
A
A
Hooper
M
M
Berns
A
A
D000818:Animals; D002460:Cell Line; D003062:Codon / Q000235:genetics; D004622:Embryo, Mammalian; D004625:Embryo, Nonmammalian; D006412:Hematopoietic Stem Cells; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051573:Proto-Oncogene Proteins c-pim-1; D011519:Proto-Oncogenes; D011995:Recombination, Genetic; D015183:Restriction Mapping
D003062:Codon; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D051573:Proto-Oncogene Proteins c-pim-1