Muzumdar Mandar Deepak, Tasic Bosiljka, Miyamichi Kazunari, Li Ling, Luo Liqun
Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
Genesis. 2007 Sep;45(9):593-605. doi: 10.1002/dvg.20335.
The Cre/loxP system has been used extensively for conditional mutagenesis in mice. Reporters of Cre activity are important for defining the spatial and temporal extent of Cre-mediated recombination. Here we describe mT/mG, a double-fluorescent Cre reporter mouse that expresses membrane-targeted tandem dimer Tomato (mT) prior to Cre-mediated excision and membrane-targeted green fluorescent protein (mG) after excision. We show that reporter expression is nearly ubiquitous, allowing visualization of fluorescent markers in live and fixed samples of all tissues examined. We further demonstrate that mG labeling is Cre-dependent, complementary to mT at single cell resolution, and distinguishable by fluorescence-activated cell sorting. Both membrane-targeted markers outline cell morphology, highlight membrane structures, and permit visualization of fine cellular processes. In addition to serving as a global Cre reporter, the mT/mG mouse may also be used as a tool for lineage tracing, transplantation studies, and analysis of cell morphology in vivo.
Cre/loxP系统已被广泛用于小鼠的条件性诱变。Cre活性报告基因对于定义Cre介导的重组的时空范围很重要。在此，我们描述了mT/mG，一种双荧光Cre报告基因小鼠，在Cre介导的切除之前表达膜靶向串联二聚体番茄红素（mT），切除后表达膜靶向绿色荧光蛋白（mG）。我们表明报告基因表达几乎无处不在，允许在所有检查组织的活样本和固定样本中可视化荧光标记。我们进一步证明mG标记是Cre依赖性的，在单细胞分辨率下与mT互补，并且可以通过荧光激活细胞分选区分。两种膜靶向标记都勾勒出细胞形态，突出膜结构，并允许可视化精细的细胞过程。除了作为全局Cre报告基因外，mT/mG小鼠还可作为谱系追踪、移植研究和体内细胞形态分析的工具。
