A global double-fluorescent cre reporter mouse
A global double-fluorescent cre reporter mouse
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DOI：
10.1002/dvg.20335
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发表时间：
2007-09-01
期刊：
影响因子：
1.5
通讯作者：
Luo, Liqun
中科院分区：
文献类型：
--
作者：
Muzumdar, Mandar Deepak;Tasic, Bosiljka;Luo, Liqun
Cre/loxP 系统已广泛用于小鼠条件诱变。 Cre 活动的报告基因对于定义 Cre 介导的重组的空间和时间范围非常重要。在这里，我们描述了 mT/mG，一种双荧光 Cre 报告小鼠，在 Cre 介导的切除之前表达膜靶向串联二聚体番茄 (mT)，在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，使得荧光标记可以在所有检查组织的活体和固定样本中可视化。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来区分。两种膜靶向标记物都勾勒出细胞形态，突出膜结构，并允许精细细胞过程的可视化。除了充当全球 Cre 报告基因外，mT/mG 小鼠还可用作谱系追踪、移植研究和体内细胞形态分析的工具。
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.