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Embryonic stem (ES) cells, which can differentiate into almost all types of cells, have been derived from the house mouse  Mus musculus , rat, rabbit, humans, and other species. Transmission of the genotype to the offspring of chimeras has been achieved only with  M. musculus  ES cells, limiting targeted mutagenesis using ES cells to this species.  Mus spretus , which exhibits many genetic polymorphisms with  M. musculus , displays dominant resistance to cancer and inflammation, making derived inbred strains very useful in positional cloning and interspecies mapping. We show here for the first time the derivation of ES cells from hybrid blastocysts, obtained by the mating of two different species, namely  Mus musculus  and  Mus spretus , and their use for the generation of chimeric mice that transmit the  Mus spretus  genotype and phenotype to the offspring. These hybrid ES cells allow the genetic manipulation of  Mus spretus , as an alternative to  Mus musculus.

Breaking the Species Barrier: Derivation of Germline‐Competent Embryonic Stem Cells from Mus spretus × C57BL/6 Hybrids