Li Wen-Long, Cheng Xuan, Tan Xiao-Hong, Zhang Ji-Shuai, Sun Yan-Song, Chen Lin, Yang Xiao
Genetic Laboratory of Development and Diseases, Institute of Biotechnology, Beijing 100071, China.
Yi Chuan Xue Bao. 2005 Sep;32(9):909-15.
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.
内皮细胞参与血管生成、血管稳态、血栓形成、炎症反应及血管壁重塑。为利用Cre-loxP系统研究内皮细胞中基因的功能,我们构建了Tie2-Cre转基因小鼠,其中Cre重组酶的表达由Tie2启动子驱动。通过基因组PCR和Southern印迹鉴定出总共6只携带Tie2-Cre转基因的奠基小鼠。整合效率为11%。为检测Cre重组酶的切割活性和组织分布,将Tie2-Cre转基因品系与携带Smad4条件等位基因的小鼠品系(Smad4(Co/Co))或报告品系ROSA26进行杂交。对Tie2-Cre; Smad4(Co/+)小鼠多个组织的DNA进行PCR检测,结果显示在所有含有内皮细胞的组织中均有Cre活性。通过lacZ染色,我们在Tie2-Cre; ROSA26双转基因胚胎中检测到了Cre转基因的泛内皮表达。因此,该小鼠品系可作为内皮细胞谱系分析和内皮细胞中条件性基因敲除的宝贵工具。