多药耐药基因1(Abcb1)敲除和人源化大鼠模型的建立  

Establishment of Abcb1 knock out rat and Abcb1 humanized rat models

作　　者：马婧[1] 陈炜[1] 张旭[1] 马元武[1] 吕丹[1] 高虹[1] 张连峰[1] 

机构地区：[1]中国医学科学院&北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,北京100021

出　　处：《中国比较医学杂志》2015年第3期1-8,共8页Chinese Journal of Comparative Medicine

基　　金：国家"重大新药创制"科技重大专项课题基于人源化模型和肝脏生物功能网络的药物肝毒性预测关键新技术(2012ZX09301001-006)

摘　　要：目的敲除大鼠多药耐药基因1(Abcb1),并在基因敲除大鼠的基础上建立Abcb1人源化大鼠模型,为Abcb1相关药物代谢和药物评价研究提供更接近人类的动物模型。方法利用大片段转基因技术和CRISPR/Cas9技术相结合,建立人源化大鼠模型,利用PCR,RT-PCR和Real-time PCR的方法进行鉴定及分析。结果将包含人源Abcb1启动子和c DNA的153 kb BAC片断转入到大鼠基因组,获得稳定表达h-Abcb1基因的大鼠,同时建立了Abcb1基因敲除大鼠。通过将两者杂交建立了Abcb1人源化大鼠模型。人源大鼠与大鼠内源Abcb1表达谱有明显的区别,人源化大鼠不仅表达h-Abcb1基因,在组织表达谱方面也与人类更接近。结论建立了Abcb1基因敲除大鼠和人源化大鼠模型,Abcb1人源化模型可作为Abcb1基因相关药物代谢研究更接近人类的动物模型。Objective To knock out the Abcbl gene of rat,and establish the Abcbl humanized rat model based on the Abcbl knock out rat. Methods The animal model was established using BAC and CRISPR/Cas9 technology,and was analyzed by PCR, RT-PCR and real-time PCR. Results Establishing a rat model expressing human Abcbl stably by transfer the 153 kb BAC containing human Abcbl promoter and cDNA into rat genome, and establishing the Abcbl knock out rat at the same time. Establishing the Abcbl humanized model by crossing these two strains together. The expression pattern of Abcbl in Abcbl humanized rat is different from the wild type rat. The Abcbl humanized model express not only the human Abcbl gene but also has similar expression pattern as human. Conclusions The Abcbl knock out rat and the Abcbl humanized rat were successfully established, and this model is close to human concerning about the drug metabolism related to Abcbl.

关 键 词：ABCB1 人源化 BAC 基因敲除大鼠 

分 类 号：R332[医药卫生—人体生理学]