Generating Tamoxifen-Inducible Cre Alleles to Investigate Myogenesis in Mice
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-ERT2 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).
Key wordsPax7 Myogenic progenitor Satellite cell Lineage tracing Tamoxifen Cre Cre-ERT2 Homologous recombination Recombineering Knock-out Knock-in
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