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血管内皮细胞参与血管形成、血管稳态维持、血栓形成、炎症和血管重建等生理和病理过程。为了便于通过Cre-LoxP系统研究相关基因在血管内皮细胞中的功能,创建了Tie2-Cre转基因小鼠,利用Tie2基因的启动子驱动Cre重组酶基因在血管内皮细胞中表达。经基因组PCR和SouthernBlot鉴定有6只小鼠在基因组上整合有Cre基因,整合率为11%。为了验证Cre重组酶的剪切活性和表达组织分布,我们将Tie2-Cre转基因小鼠分别与Smad4条件基因打靶小鼠和报告小鼠ROSA26交配。Tie2-Cre;Smad4Co/+小鼠的多个组织的基因组DNA的PCR结果显示,Cre重组酶在所有包含血管内皮细胞的组织中表达并能介导LoxP间的重组。Tie2-Cre;ROSA26双转基因胚胎LacZ染色结果显示,Cre重组酶在所有被检测组织的血管内皮细胞中特异性表达。因此,Tie2-Cre转基因小鼠可作血管内皮细胞谱系分析和在血管内皮细胞进行条件基因打靶的理想工具小鼠。
Vascular endothelial cells are involved in physiological and pathological processes such as angiogenesis, maintenance of vascular homeostasis, thrombosis, inflammation and revascularization. In order to facilitate the study of the function of related genes in vascular endothelial cells by the Cre-LoxP system, Tie2-Cre transgenic mice were created and the Cre recombinase gene was driven by the promoter of Tie2 gene to be expressed in vascular endothelial cells. Six genomic DNAs were identified by genomic PCR and SouthernBlot. Cre gene was integrated into the genome and the integration rate was 11%. To verify the cleavage activity and tissue distribution of Cre recombinase, we crossed Tie2-Cre transgenic mice with Smad4 -conjugated and ROSA26-challenged mice, respectively. PCR results of genomic DNA from multiple tissues of Tie2-Cre; Smad4Co / + mice revealed that Cre recombinase is expressed in all tissues containing vascular endothelial cells and mediates recombination between LoxP. Tie2-Cre; LacZ staining of ROSA26 double transgenic embryos showed that Cre recombinase was specifically expressed in vascular endothelial cells of all tested tissues. Therefore, Tie2-Cre transgenic mice can be used as an ideal tool for vascular endothelial cell lineage analysis and target gene expression in vascular endothelial cells.