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 | Zendy

Masahiko Taniguchi | Zendy; Makoto Sanbo | Zendy; Satoshi Watanabe | Zendy; I. Naruse | Zendy; Masahiro Mishina | Zendy; Takeshi Yagi | Zendy

Open Access

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

Author(s) -

Masahiko Taniguchi,

Makoto Sanbo,

Satoshi Watanabe,

I. Naruse,

Masahiro Mishina,

Takeshi Yagi

Publication year - 1998

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/26.2.679

Subject(s) - puromycin , biology , gene , gene targeting , microbiology and biotechnology , mutant , marker gene , genetics , protein biosynthesis

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.

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