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目的:检测Cre重组酶在心肌细胞特异性Cre重组酶转基因小鼠(α-MHC-Cre)中的组织分布及其在体内介导基因重组的作用。方法:将α-MHC-Cre转基因小鼠与ROSA26报告小鼠交配,利用LacZ染色对双转基因阳性子代小鼠进行检测。然后,将α-MHC-Cre小鼠与Sm ad4条件基因打靶小鼠交配,利用PCR和Southern杂交对Cre重组酶介导重组的组织特异性进行检测。结果:LacZ染色表明,Cre重组酶只在心肌细胞中特异性表达并介导ROSA位点LoxP序列间的重组。PCR和Southern结果显示Cre重组酶只在心肌细胞中特异地剔除了Sm ad4基因,进一步验证了Cre重组酶在心肌细胞中发挥介导LoxP位点重组的作用。结论:α-MHC-Cre转基因小鼠具有良好的组织特异性,只在心肌细胞中表达Cre重组酶,并能在体内成功地介导心肌细胞基因组上LoxP位点间的重组,是一种理想的研制心肌细胞特异性基因剔除小鼠的工具小鼠。
AIM: To investigate the tissue distribution of Cre recombinase in cardiomyocyte-specific Cre recombinase transgenic mice (α-MHC-Cre) and its role in mediating gene recombination in vivo. METHODS: Alpha-MHC-Cre transgenic mice were mated with ROSA26 reporter mice, and double transgenic positive offspring mice were detected by LacZ staining. Then, α-MHC-Cre mice were crossed with Sm ad4 conditional gene-targeted mice, and the tissue specificity of Cre recombinase-mediated recombination was detected by PCR and Southern hybridization. Results: LacZ staining showed that Cre recombinase was only expressed specifically in cardiomyocytes and mediated the recombination between the LoxP sequences at the ROSA site. PCR and Southern results showed that Cre recombinase only specifically knocked down the Sm ad4 gene in cardiomyocytes, further confirming the role of Cre recombinase in cardiomyocyte mediated LoxP site recombination. CONCLUSIONS: Alpha-MHC-Cre transgenic mice have a good tissue specificity and are ideal only to express Cre recombinase in cardiomyocytes and to successfully mediate recombination of LoxP sites on the cardiomyocyte genome in vivo Development of cardiomyocyte-specific knockout mice in a tool mouse.