Southard Sheryl, Low SiewHui, Li Lydia, Rozo Michelle, Harvey Tyler, Fan Chen-Ming, Lepper Christoph
Department of Embryology, Carnegie Institution for Science, Baltimore, Maryland.
Genesis. 2014 Aug;52(8):759-70. doi: 10.1002/dvg.22792. Epub 2014 Jun 3.
We report the generation of five mouse strains with the tamoxifen-inducible Cre (Cre-ER(T) (2) ; CE) gene cassette knocked into the endogenous loci of Pax3, Myod1, Myog, Myf6, and Myl1, collectively as a resource for the skeletal muscle research community. We characterized these CE strains using the Cre reporter mice, R26R(L) (acZ) , during embryogenesis and show that they direct tightly controlled tamoxifen-inducible reporter expression within the expected cell lineage determined by each myogenic gene. We also examined a few selected adult skeletal muscle groups for tamoxifen-inducible reporter expression. None of these new CE alleles direct reporter expression in the cardiac muscle. All these alleles follow the same knock-in strategy by replacing the first exon of each gene with the CE cassette, rendering them null alleles of the endogenous gene. Advantages and disadvantages of this design are discussed. Although we describe potential immediate use of these strains, their utility likely extends beyond foreseeable questions in skeletal muscle biology.
我们报告了五种小鼠品系的产生,其携带的他莫昔芬诱导型Cre(Cre-ER(T) (2) ; CE)基因盒被敲入Pax3、Myod1、Myog、Myf6和Myl1的内源性位点,共同作为骨骼肌研究群体的一种资源。我们在胚胎发育过程中使用Cre报告基因小鼠R26R(L) (acZ)对这些CE品系进行了表征,结果表明它们在由每个生肌基因确定的预期细胞谱系内指导严格控制的他莫昔芬诱导型报告基因表达。我们还检查了一些选定的成年骨骼肌组中他莫昔芬诱导型报告基因的表达情况。这些新的CE等位基因均未在心肌中指导报告基因表达。所有这些等位基因都采用相同的敲入策略,即用CE盒替换每个基因的第一个外显子,使其成为内源性基因的无效等位基因。本文讨论了这种设计的优缺点。虽然我们描述了这些品系可能的直接用途,但其效用可能超出骨骼肌生物学中可预见的问题。