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

Daniel Metzger1, Pierre Chambon1

  • 1Institut de Génétique et de Biologie Moléculaire et Cellulaire, Université de Strasbourg, and Collège de France, Illkirch, France.

Current Protocols in Mouse Biology
|June 13, 2015
PubMed

Insights

This study details a tamoxifen-inducible Cre-ER(T2) system for precise genetic mutations in mouse skin cells. The method enables controlled gene editing in adult mice for disease modeling.

Area of Science:

  • Genetics
  • Molecular Biology
  • Dermatology

Background:

  • Ligand-activated Cre recombinases enable targeted somatic mutagenesis.
  • Cre-ER(T2) recombinase activity can be controlled by tamoxifen.

Purpose of the Study:

  • To describe a protocol for temporally controlled, targeted somatic mutagenesis in mouse epidermal keratinocytes.
  • To demonstrate the utility of K14-Cre-ER(T2) transgenic mice for inducing mutations in adult mice.

Main Methods:

  • Utilizing K14-Cre-ER(T2) transgenic mice expressing tamoxifen-activatable Cre recombinase.
  • Administering low-dose tamoxifen (100-µg daily for 5 days) to induce Cre-ER(T2) activity.
  • Inducing site-specific somatic mutations in epidermal keratinocytes of adult mice.

Main Results:

  • Highly reproducible generation of site-directed somatic mutations in mouse epidermal keratinocytes.
  • Successful application of the technique in creating mouse models for human diseases.
  • Demonstrated efficacy of low-dose tamoxifen for controlled genetic manipulation.

Conclusions:

  • The described protocol provides a reliable method for temporal control of targeted somatic mutations in mouse epidermis.
  • This technique facilitates the generation of genetically modified mouse models for studying human diseases.
  • Ligand-inducible Cre-ER(T2) systems offer precise tools for somatic mutagenesis in specific cell types.
Keywords:
Cre-ERT2keratinocytesloxPskintamoxifen

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