Production of chimaeric mice containing embryonic stem (ES) cells carrying a homoeobox Hox 1.1 allele mutated by homologous recombination.
Several mouse gene families related to Drosophila developmental control genes and containing a homoeobox, a paired box or a finger domain, have been cloned and structurally analysed. On the basis of structural similarities to the Drosophila genes and of their spatially and temporally restricted expression patterns during mouse embryogenesis, it has been proposed that these mammalian genes also are involved in the control of development. To elucidate the function of homoeobox genes by genetic means, mouse mutants must be generated. We have developed a technique for mutagenesis in vivo and have used it to mutate the homoeobox Hox 1.1 gene. In vivo mutagenesis was achieved through homologous recombination between an endogenous Hox 1.1 allele and a microinjected mutated gene in pluripotent embryonic stem (ES) cells. Mutant cells were identified by means of the polymerase chain reaction (PCR) and mutant clones were used to generate chimaeric mice. Because the homologous recombination event is formally a gene conversion event and no selection is required to screen for cells carrying the mutated allele, in vivo mutagenesis allows specific alterations in the target sequence to be made without the introduction of any other sequences.
Zimmer
A
A
Max-Planck Institut für biophysikalische Chemie, Abt. Molekulare Zellbiologie, Göttingen, FRG.
Gruss
P
P
D000818:Animals; D015139:Blotting, Southern; D002678:Chimera; D003001:Cloning, Molecular; D005801:Genes, Homeobox; D051379:Mice; D009154:Mutation; D013234:Stem Cells; D014162:Transfection