Novak A, Guo C, Yang W, Nagy A, Lobe C G
Cancer Research Division, Sunnybrook and Women's College Health Science Centre, Toronto, Ontario, Canada.
Genesis. 2000 Nov-Dec;28(3-4):147-55.
The Cre/loxP system has become an important tool in designing postintegrational switch mechanisms for transgenes in mice. The power and spectrum of application of this system depends on transgenic mouse lines that provide Cre recombinase activity with a defined cell type-, tissue-, or developmental stage-specificity. We have developed a novel mouse line that acts as a Cre reporter. The mice, designated Z/EG (lacZ/EGFP), express lacZ throughout embryonic development and adult stages. Cre excision, however, removes the lacZ gene, which activates expression of the second reporter, enhanced green fluorescent protein. We have found that the double-reporter Z/EG line is able to indicate the occurrence of Cre excision from early embryonic to adult lineages. The advantage of the Z/EG line is that Cre-mediated excision can be monitored in live samples and that live cells with Cre-mediated excision can be isolated using a single-step FACS. 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系统已成为设计小鼠转基因整合后开关机制的重要工具。该系统的应用能力和范围取决于能在特定细胞类型、组织或发育阶段提供Cre重组酶活性的转基因小鼠品系。我们开发了一种新型的作为Cre报告基因的小鼠品系。这些小鼠被命名为Z/EG(lacZ/EGFP),在整个胚胎发育和成年阶段均表达lacZ。然而,Cre切除会去除lacZ基因,从而激活第二个报告基因——增强型绿色荧光蛋白的表达。我们发现双报告基因Z/EG品系能够指示从早期胚胎到成年谱系中Cre切除的发生情况。Z/EG品系的优势在于可以在活样本中监测Cre介导的切除,并且可以通过单步荧光激活细胞分选术(FACS)分离出具有Cre介导切除的活细胞。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。