Center for Muscle Biology, University of Kentucky, Lexington, KY, 40536, USA.
Skelet Muscle. 2012 May 7;2(1):8. doi: 10.1186/2044-5040-2-8.
The use of the Cre/loxP system for gene targeting has been proven to be a powerful tool for understanding gene function. The purpose of this study was to create and characterize an inducible, skeletal muscle-specific Cre transgenic mouse strain.
To achieve skeletal muscle-specific expression, the human α-skeletal actin promoter was used to drive expression of a chimeric Cre recombinase containing two mutated estrogen receptor ligand-binding domains.
Western blot analysis, PCR and β-galactosidase staining confirmed that Cre-mediated recombination was restricted to limb and craniofacial skeletal muscles only after tamoxifen administration.
A transgenic mouse was created that allows inducible, gene targeting of floxed genes in adult skeletal muscle of different developmental origins. This new mouse will be of great utility to the skeletal muscle community.
Cre/loxP 系统在基因靶向中的应用已被证明是一种研究基因功能的强大工具。本研究的目的是构建和鉴定一种可诱导的、骨骼肌特异性 Cre 转基因小鼠品系。
为实现骨骼肌特异性表达,使用人α-骨骼肌肌动蛋白启动子驱动包含两个突变雌激素受体配体结合域的嵌合 Cre 重组酶的表达。
Western blot 分析、PCR 和β-半乳糖苷酶染色证实,只有在给予他莫昔芬后,Cre 介导的重组才局限于四肢和颅面骨骼肌。
构建了一种转基因小鼠,可在不同发育来源的成年骨骼肌中诱导、靶向 floxed 基因。这种新型小鼠将对骨骼肌研究领域具有重要的应用价值。