Use of Chemical Mutagenesis in Mouse Embryonic Stem Cells

The chemical mutagenesis of mouse embryonic stem (ES) cells is an approach complementary to chemical mutagenesis of spermatogonia in whole animals. It has great potential to contribute significantly to the generation of a comprehensive collection of multiple alleles for most mammalian genes and to facilitate the progression from gene sequence to gene functional analyses. The general strategy includes the treatment of ES cells with a chemical mutagen and the isolation of individual mutagenized clones that are then cultured in duplicate. Whereas one set of samples is cryopreserved for archiving and subsequent generation of germline chimeras by means of blastocyst injection, the second set is used for genotypeor phenotype-driven screens to identify mutant alleles.