Fiche publication
Date publication
décembre 1997
Journal
Proceedings of the National Academy of Sciences of the United States of America
Auteurs
Membres identifiés du Cancéropôle Est :
Pr CHAMBON Pierre
,
Dr METZGER Daniel
,
Dr VONESCH Jean-Luc
Tous les auteurs :
Brocard J, Warot X, Wendling O, Messaddeq N, Vonesch JL, Chambon P, Metzger D
Lien Pubmed
Résumé
The efficient introduction of somatic mutations in a given gene, at a given time, in a specific cell type will facilitate studies of gene function and the generation of animal models for human diseases. We have shown previously that conditional recombination-excision between two loxP sites can be achieved in mice by using the Cre recombinase fused to a mutated ligand binding domain of the human estrogen receptor (Cre-ERT), which binds tamoxifen but not estrogens. DNA excision was induced in a number of tissues after administration of tamoxifen to transgenic mice expressing Cre-ERT under the control of the cytomegalovirus promoter. However, the efficiency of excision varied between tissues, and the highest level ( approximately 40%) was obtained in the skin. To determine the efficiency of excision mediated by Cre-ERT in a given cell type, we have now crossed Cre-ERT-expressing mice with reporter mice in which expression of Escherichia coli beta-galactosidase can be induced through Cre-mediated recombination. The efficiency and kinetics of this recombination were analyzed at the cellular level in the epidermis of 6- to 8-week-old double transgenic mice. We show that site-specific excision occurred within a few days of tamoxifen treatment in essentially all epidermis cells expressing Cre-ERT. These results indicate that cell-specific expression of Cre-ERT in transgenic mice can be used for efficient tamoxifen-dependent, Cre-mediated recombination at loci containing loxP sites to generate site-specific somatic mutations in a spatio-temporally controlled manner.
Mots clés
Animals, Epidermis, metabolism, Gene Targeting, Humans, Integrases, genetics, Mice, Mice, Transgenic, Mutagenesis, Site-Directed, Receptors, Estrogen, genetics, Recombinant Fusion Proteins, genetics, Recombination, Genetic, Tamoxifen, metabolism, Viral Proteins
Référence
Proc. Natl. Acad. Sci. U.S.A.. 1997 Dec;94(26):14559-63
