Gene inactivation has become the gold standard for determining gene function in the mouse. Many genes inactivated in the germ line cause early lethality that precludes phenotypic assessment at a later time point. Conditional gene inactivation using Cre recombinase expressed via a tissue specific promoter/enhancer allows phenotypic analyses of selected tissues, but lacks temporal control. Recent development of the tamoxifen-inducible Cre-ER T2 offers both cell type-specific and temporal control of conditional gene inactivation. As an example, we describe the design and step-wise construction of a Cre-ER T2 knock-in allele at the Pax7 locus using the recombineering method - Pax7 is selectively expressed in embryonic muscle progenitors and adult muscle stem cells. The resulting Pax7- C re- E R T2 (Pax7 CE ) allele has been successfully applied to embryos and adults for tamoxifen-regulated myogenic lineage tracing and gene inactivation (Nature 460:627-631, 2009; Genesis 48:424-436, 2010).
CITATION STYLE
Lepper, C., & Fan, C. M. (2012). Generating tamoxifen-inducible cre alleles to investigate myogenesis in mice. Methods in Molecular Biology, 798, 297–308. https://doi.org/10.1007/978-1-61779-343-1_17
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