血管内皮细胞特异表达Cre重组酶转基因小鼠的建立(英文)  

Endothelial Cell-Specific Expression of Cre Recombinase in Transgenic Mice

作　　者：李文龙[1] 程萱[1] 谭晓红[1] 张继帅[1] 孙岩松[2] 陈林[3] 杨晓[1] 

机构地区：[1]军事医学科学院生物工程研究所发育和疾病遗传学研究室,北京100071 [2]军事医学科学院实验动物中心,北京100071 [3]第三军医大学附属大坪医院全军战创伤中心实验室,重庆400042

出　　处：《Acta Genetica Sinica》2005年第9期909-915,共7页

基　　金：国家杰出青年基金(编号:30025028,30128013);国家自然科学基金(编号:30400252,30370318)~~

摘　　要：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。Endothelial ceiis participate in angiogenesis, vascuiar 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.

关 键 词：TIE2 血管内皮细胞 CRE-LOXP 转基因小鼠 CRE重组酶 特异表达 基因组DNA CRE-LOXP Southern 组织分布 

分 类 号：R363[医药卫生—病理学]