Generating Targeted Gene Alterations

The most powerful technology for genetic manipulation in mammals gene targeting was developed in mice, and depends on the ability of mammalian DNA, when added to cells in culture, to recombine homologously with identical DNA sequences in the genome and to replace them. In mice, this can be achieved by the use of embryonic stem (ES) cells. However to date, there are no fully validated ES cell lines in domestic animals. Nuclear transfer from nonpluripotent cell lines provides an alternative to the ES cell route for the introduction of targeted gene alterations into the germ line. Successful production of cloned pigs for xenotransplantation resulting from the combination of somatic nuclear transfer with specific modification (knock out) of fetal fibroblasts has been reported.[14]

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