Fiche publication
Date publication
novembre 2000
Journal
Experimental cell research
Auteurs
Membres identifiés du Cancéropôle Est :
Pr CHAMBON Pierre
,
Dr METZGER Daniel
Tous les auteurs :
Chiba H, Chambon P, Metzger D
Lien Pubmed
Résumé
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.
Mots clés
Carcinoma, Embryonal, Doxycycline, pharmacology, Gene Expression, drug effects, Genetic Engineering, Humans, Integrases, genetics, Mutagenesis, Site-Directed, Receptors, Estrogen, genetics, Repressor Proteins, genetics, Retinoids, pharmacology, Tamoxifen, pharmacology, Tetracycline, pharmacology, Tumor Cells, Cultured, Viral Proteins
Référence
Exp. Cell Res.. 2000 Nov;260(2):334-9
