Generation of PDE4 knockout mice by gene targeting.

摘要: The development of gene-targeting techniques has ushered in a new era mouse genetics. Two discoveries have been instrumental: the finding that an exogenous DNA introduced mammalian cells can recombine with homologous chromosomal sequences, process known as gene targeting, and revelation cultured embryonic stem (ES) when injected into early stage embryos contribute to produce germ-line chimeras. On basis these seminal findings, targeting by recombination ES vitro established powerful means altering specific loci genome. As result, function be studied vivo. By applying this technology, targeted disruption PDE4 alleles is created and, subsequently, mutant are blastocysts returned pseudopregnant foster mothers chimeric pups. In chapter, we describe basic protocols used generate knockout mice.