利用基因打靶技术构建血管内皮特异性Stk24/25双基因敲除小鼠

目的:构建Stk24/Stk25双基因敲除小鼠模型，并对其基因型及敲除效率进行测定。方法:使用基因打靶技术构建基因敲除小鼠，利用PCR及凝胶电泳技术对亲代及子代小鼠进行基因型鉴定；并利用ＲT-qPCＲ测定目的基因在小脑血管内皮细胞中mＲNA转录水平的表达情况。结果:通过PCR技术成功检测出小鼠子代基因型Cdh5cre+/Stk24fl/fl/Stk25fl/fl纯合子小鼠，经鉴定血管内皮基因敲除Stk24、Stk25后表达量分别下降了27%（t=2.874，P=0.0207）和30%（t=17.21，P<0.001）。结论:采用基因打靶技术成功构建了Stk24/Stk25双基因敲除小鼠。

Establishment of a mouse model with endothelial-special Stk24/25 knockout with gene targeting technique

Objective:To generate endothelial specific Stk24 and Stk25 gene-knockout mice using gene targeting technology，and identify the genotype and the knockout efficiency of Stk24/25 mouse strains. Methods: Gene knockout mice were established with gene targeting technology. The genotypes of mouse were identified by PCR and gel electrophoresis. The expression of target mRNA in cerebellum endothelial cells was detected by RT-qPCR. Results: The genotypes of the mice were successfully identified by PCR，including Cdh5cre+-Stk24fl/fl-Stk25fl/fl mice. The expression level of Stk24，Stk25 mRNA in the cerebellum endothelial cells of Stk24/Stk25 gene knockout mice significantly reduced by 27%（t=2.874，P=0.0207） and 30%（t=17.21，P<0.001），respectively. Conclusion: The mouse models with endothelial specific Stk24/25 gene knockout are successfully established by using gene targeting technique.