Generation of Spatio-Temporally Controlled Targeted Somatic Mutations in the Mouse.

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Date publication

janvier 2011

Auteurs

Membres identifiés du Cancéropôle Est :
Pr CHAMBON Pierre, Dr METZGER Daniel


Tous les auteurs :
Metzger D, Chambon P

Résumé

The generation of ligand-activated site-specific Cre recombinases has led to the development of cell type-specific temporally controlled targeted somatic mutagenesis in the mouse. We illustrate this technique using K14-Cre-ER(T2) transgenic mice that express the tamoxifen (tam)-activatable Cre-ER(T2) recombinase in epidermal basal keratinocytes to induce mutations in epidermal keratinocytes of adult mice. Our highly reproducible technique, based on induction of Cre-ER(T2) recombinase activity by tamoxifen administration at low doses (once daily 100-microg intraperitoneal injection for 5 days), has allowed the generation of site-directed somatic mutations of numerous genes in mouse epidermal keratinocytes, and several mouse models of human diseases. The present step-by-step protocol describes how to introduce temporally controlled targeted mutations in epidermal keratinocytes of adult mice. Curr. Protoc. Mouse Biol. 1:55-70. (c) 2011 by John Wiley & Sons, Inc.

Référence

Curr Protoc Mouse Biol. 2011 Mar 1;1(1):55-70