[A new universal gene replacement method for generating a desired mutant mouse].
Site-directed gene replacement has been accomplished using two steps of gene targeting, which is called universal gene replacement. The previously described universal gene replacement method with the "neo-tk" cassette exhibited a very low efficiency in the second "replacement" step. The neo-tk tagged allele obtained from the first step was never transmitted to offspring through the germ cells of chimeric mice. We therefore developed a new system using the Eco-gpt gene instead of the HSV-tk gene. With this system, gene replacement for the p53 gene was thus consistently accomplished and generated mutant mouse lines with site-directed mutations. Further, all manipulated alleles, including the neo-gpt tagged one, were successfully transmitted to offspring.
Itoh
K
K
Department of Molecular and Cellular Biology, Kyushu University, Fukuoka.
D000818:Animals; D002678:Chimera; D018390:Gene Targeting; D016158:Genes, p53; D005821:Genetic Techniques; D051379:Mice; D011995:Recombination, Genetic