Lobe C G, Koop K E, Kreppner W, Lomeli H, Gertsenstein M, Nagy A
Cancer Research Division, Sunnybrook Health Science Centre, 2075 Bayview Avenue, Toronto, Ontario, M4N 3M5, Canada.
Dev Biol. 1999 Apr 15;208(2):281-92. doi: 10.1006/dbio.1999.9209.
The Cre/loxP site-specific recombination system combined with embryonic stem cell-mediated technologies has greatly expanded our capability to address normal and disease development in mammals using genetic approaches. The success of this emerging technology hinges on the production of Cre-expressing transgenic lines that provide cell type-, tissue-, or developmental stage-specific recombination between loxP sites placed in the genome. Here we describe and characterize the production of a double-reporter mouse line that provides a convenient and reliable readout of Cre recombinase activity. Throughout all embryonic and adult stages, the transgenic animal expresses the lacZ reporter gene before Cre-mediated excision occurs. Cre excision, however, removes the lacZ gene, allowing expression of the second reporter, the human alkaline phosphatase gene. This double-reporter transgenic line is able to indicate the occurrence of Cre excision in an extremely widespread manner from early embryonic to adult lineages. It will be a valuable reagent for the increasing number of investigators taking advantage of the powerful tools provided by the Cre/loxP site-specific recombinase system.
Cre/loxP位点特异性重组系统与胚胎干细胞介导技术相结合,极大地扩展了我们运用遗传学方法研究哺乳动物正常发育和疾病发生的能力。这项新兴技术的成功取决于表达Cre的转基因品系的产生,这些品系能在基因组中loxP位点之间实现细胞类型、组织或发育阶段特异性重组。在此,我们描述并鉴定了一种双报告基因小鼠品系的产生,该品系能方便、可靠地读出Cre重组酶活性。在所有胚胎和成年阶段,转基因动物在Cre介导的切除发生之前表达lacZ报告基因。然而,Cre切除会去除lacZ基因,从而使第二个报告基因——人碱性磷酸酶基因得以表达。这种双报告基因转基因品系能够以极其广泛的方式指示从早期胚胎到成年谱系中Cre切除的发生情况。对于越来越多利用Cre/loxP位点特异性重组酶系统提供的强大工具的研究人员来说,它将是一种有价值的试剂。