Cre重组酶在转基因小鼠内皮细胞中的特异性表达。

Wen-Long Li, Xuan Cheng, Xiao-Hong Tan, Ji-Shuai Zhang, Yan-Song Sun, Lin Chen, Xiao Yang
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摘要

内皮细胞参与血管生成、血管稳态、血栓形成、炎症和血管壁重塑。为了利用Cre- loxp系统研究基因在内皮细胞中的功能,我们培育了由Tie2启动子驱动Cre重组酶表达的Tie2-Cre转基因小鼠。采用基因组PCR和Southern blot方法鉴定了6只携带Tie2-Cre基因的创始小鼠。集成效率为11%。为了检测Cre重组酶的切除活性和组织分布,将Tie2-Cre转基因系与携带Smad4条件等位基因的小鼠品系(Smad4(Co/Co))或报告系ROSA26杂交。Tie2-Cre多组织DNA的PCR分析Smad4(Co/+)小鼠在含内皮细胞的所有组织中均显示Cre活性。我们在Tie2-Cre中检测了Cre转基因的泛内皮表达;ROSA26双转基因胚胎lacZ染色。因此,该小鼠系可以作为内皮细胞谱系分析和内皮细胞条件基因消融的有价值的工具。
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Endothelial cell-specific expression of Cre recombinase in transgenic mice.

Endothelial cells participate in angiogenesis, vascular homeostasis, thrombosis, inflammation and vascular wall remodeling. To study the function of genes in endothelial cells using Cre-loxP system, we generated Tie2-Cre transgenic mice, in which expression of Cre recombinase is driven by Tie2 promoter. Total six founder mice carrying the Tie2-Cre transgene were identified by genomic PCR and Southern blot. The integration efficiency is 11%. In order to test the excision activity and tissue distribution of the Cre recombinase, the Tie2-Cre transgenic line was crossed with the mouse strain carrying the Smad4 conditional alleles (Smad4(Co/Co)) or the reporter line ROSA26. PCR of multiple tissue DNA from Tie2-Cre; Smad4(Co/+) mice revealed the Cre activity in all tissues containing endothelial cells. We detected pan-endothelial expression of the Cre transgene in Tie2-Cre; ROSA26 double transgenic embryos by lacZ staining. Therefore, this mouse line may serve as a valuable tool for endothelial cell lineage analyses and conditional gene ablation in endothelial cells.

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