F9 embryonal carcinoma cells engineered for tamoxifen-dependent Cre-mediated site-directed mutagenesis and doxycycline-inducible gene expression.
The study of gene functions in complex genetic environments such as mammalian cells would greatly benefit from systems allowing a tight control of gene expression. The tetracycline-inducible gene expression system and the site-specific Cre/loxP recombination system have gained increasing popularity for conditional expression and gene disruption. To facilitate the analysis of gene functions in a cell autonomous system, we have established an F9 murine embryonal carcinoma cell line, constitutively expressing both the doxycycline-controlled transactivator rtTA and the tamoxifen-dependent Cre recombinase Cre-ER(T). The expression of a reporter gene placed under the control of tetracycline operators was induced about 1000-fold by doxycycline, and tamoxifen-induced excision of a loxP-flanked DNA segment occurred in all cells. This genetically engineered cell line, which allows, upon simple ligand addition, sophisticated genetic manipulations, such as sequential inactivation of loxP-flanked genes, and tightly controlled reexpression of their cDNAs, should be a valuable tool for studying mammalian gene functions.
Chiba
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H
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch-Cedex, 67404, France.
Chambon
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P
Metzger
D
D
D018236:Carcinoma, Embryonal; D004318:Doxycycline / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D005818:Genetic Engineering; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011960:Receptors, Estrogen / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D012176:Retinoids / Q000494:pharmacology; D013629:Tamoxifen / Q000494:pharmacology*; D013752:Tetracycline / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014764:Viral Proteins
D011960:Receptors, Estrogen; D012097:Repressor Proteins; D012176:Retinoids; D014764:Viral Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D013629:Tamoxifen; C045073:Cre recombinase; D019426:Integrases; D013752:Tetracycline; D004318:Doxycycline