Efficient production of Cre-mediated site-directed recombinants through the utilization of the puromycin resistance gene, pac: a transient gene-integration marker for ES cells.
Gene targeting in embryonic stem (ES) cells is a powerful tool for generating mice carrying specifically designed mutations in the germline. Puromycin can completely kill ES cells within 24 to 48 h whereas G418 and hygromycin cannot. We have, therefore, proposed that the puromycin N-acetyltransferase ( pac ) gene, may be utilized as a transient gene-integration marker. Using a circular expression vector of cre and pac genes, Cre-mediated mutant cells were effectively enriched by pulse treatment of puromycin without stable integration of their genes. We have thus demonstrated the first application of pac as a transient gene-integration marker for ES cells.
Taniguchi
M
M
Laboratory of Neurobiology and Behavioral Genetics, National Institute for Physiological Sciences, Myodaiji, Okazaki 444, Japan.
Sanbo
M
M
Watanabe
S
S
Naruse
I
I
Mishina
M
M
Yagi
T
T
D000123:Acetyltransferases / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D015870:Gene Expression; D018390:Gene Targeting; D005819:Genetic Markers; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011691:Puromycin / Q000494:pharmacology; D011995:Recombination, Genetic; D013234:Stem Cells / Q000378:metabolism*; D014764:Viral Proteins
D005819:Genetic Markers; D014764:Viral Proteins; D011691:Puromycin; D000123:Acetyltransferases; C045336:puromycin N-acetyltransferase; C045073:Cre recombinase; D019426:Integrases