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Transgenic mice for the establishment of histidinol‐resistant embryonic fibroblast feeder layers
Author(s) -
Tucker Rebecca M.,
Burke David T.
Publication year - 1996
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.10.14.9002557
Subject(s) - embryonic stem cell , transgene , genetically modified mouse , microbiology and biotechnology , fibroblast , biology , genetics , gene , cell culture
Gene targeting in mouse embryonic stem cells generates mutations by replacing an endogenous chromosomal region with a copy disrupted by a selectable genetic marker. The most commonly used selectable marker is the bacterial neo r gene, which confers resistance in mammalian cells to the antibiotic G418. Use of an alternative selectable marker, the Salmonella typhimurium gene huD, should provide expanded applications for gene targeting. The hisD gene encodes the protein histidinol dehydrogenase, which catalyzes the conversion of histidinol to the amino acid histidine. Histidinol is toxic to mammalian cells, while histidine is an essential mammalian amino acid. Consequently, growth selection in cultures with media containing histidinol in place of histidine occurs by both histidine starvation and histidinol poisoning. The hisD selection is being tested for potential use in gene targeting experiments with mouse embryonic stem (ES) cells. Currently, most successful gene targeting experiments use primary embryonic fibroblast feeder layers, which assist in the maintenance of the pluripotential state of the embryonic stem cells. To support ES cell stability under histidinol selection, mice transgenic for the S. typhimurium hisD gene have been produced and used to generate embryonic fibroblast feeder cells. The transgenic embryonic fibroblasts survive under a wide range of histidinol‐ containing growth conditions and support growth of ES cell cultures.—Tucker, R. M., Burke, D. T. Transgenic mice for the establishment of histidinol‐ resistant embryonic fibroblast feeder layers. FASEB J. 10, 1641‐1645 (1996)