Related Experiment Videos
Spatio-temporally controlled site-specific somatic mutagenesis in the mouse
J Brocard1, X Warot, O Wendling
1Centre National de la Recherche Scientifique, Institut National de la Santé et de la Recherche Médicale, Université Louis Pasteur, Collège de France, Strasbourg, France.
Summary
Conditional Cre-ERT recombination enables efficient, tamoxifen-inducible gene modification in specific cell types. This method allows for precise somatic mutations in animal models, advancing gene function studies and disease modeling.
Area of Science:
- Genetics
- Molecular Biology
- Developmental Biology
Background:
- Conditional recombination systems are crucial for studying gene function and creating disease models.
- Cre recombinase fused to a mutated estrogen receptor ligand-binding domain (Cre-ERT) allows tamoxifen-inducible recombination.
- Previous studies showed variable excision efficiency in different tissues.
Purpose of the Study:
- To determine the efficiency and kinetics of Cre-ERT-mediated recombination at the cellular level.
- To validate the use of Cre-ERT for spatio-temporally controlled somatic mutations.
Main Methods:
- Generation of double transgenic mice by crossing Cre-ERT expressing mice with beta-galactosidase reporter mice.
- Administration of tamoxifen to induce Cre-mediated recombination.
- Cellular-level analysis of recombination efficiency and kinetics in the epidermis.
Main Results:
- Tamoxifen treatment induced site-specific DNA excision within days.
- Essentially all epidermis cells expressing Cre-ERT showed recombination.
- High efficiency of tamoxifen-dependent, Cre-mediated recombination was demonstrated.
Conclusions:
- Cell-specific Cre-ERT expression enables efficient, tamoxifen-inducible recombination.
- This system allows for the generation of site-specific somatic mutations in a controlled manner.
- Facilitates gene function studies and the development of animal models for human diseases.