利用CRISPR/Cas9技术建立NOD/SCID/IL2Rγ^(-/-)免疫缺陷小鼠（英文）  

Generation of a new strain of NOD/SCID/IL2Rγ^(-/-) mice with targeted disruption of Prkdc and IL2Rγ genes using CRISPR/Cas9 system

作　　者：刘亚宸 陈渠 杨兴龙 汤清双 姚开泰[1] 徐洋[1] LIU Yachen;CHEN Qu;YANG Xinglong;TANG Qingshuang;YAO Kaitai;XU Yang(Guangdong Provincial Key Laboratory of Tumor Immunotherapy, Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China)

机构地区：[1]南方医科大学肿瘤研究所,广东广州510515

出　　处：《南方医科大学学报》2018年第6期639-646,共8页Journal of Southern Medical University

基　　金：Supported by Key Research Projects of National Natural Science Foundation of China(81430032,U1601222);by Special Funds for Frontier and Key Technology Innovation of Guangdong Province(2015B020225004);Major Basic Research Development Project of Natural Science Foundation of Guangdong Province~~

摘　　要：目的 NOD/SCID/IL2Rγ-(/-NSG)小鼠品系是最严重的免疫缺陷品系,广泛应用于异种移植的研究。然而,SCID突变是Prkdc基因的一个自发性突变,会导致T细胞发育阻滞后渗漏而且很难进行基因型鉴定。因此,开发一个敲除Prkdc和IL2Rγ基因的新品系NSG小鼠非常重要。方法使用CRISPR/Cas9系统实现Prkdc和IL2Rγ基因的靶向敲除。将基因敲除小鼠与NOD背景小鼠杂交,我们产生了新品系NOD/SCID/IL2Rγ-(/-NSG)小鼠,称为cNSG(中国NSG)小鼠。结果 cNSG小鼠完全缺乏T细胞,B细胞以及NK细胞而且表达NOD突变。体内成瘤实验证实cNSG鼠是一种比免疫缺陷裸鼠更有效的异种移植模型。结论 cNSG小鼠与Jackson实验室的NSG鼠的免疫缺陷一致,有望成为中国生物医学研究的重要异种移植模型。Objective The NOD/SCID/IL2 Rγ-/-（NSG） mouse strain is the most widely used immunodeficient strain for xenograft transplantation. However, the existing SCID mutation is a spontaneous mutation of the Prkdc gene, which leads to leaky T cell developmental block and difficulty in genotyping. It is therefore important to develop a new strain of NSG mice with targeted disruption of Prkdc and IL2 Rγ genes. Methods Targeted disruption of Prkdc and IL2 Rγ genes was achieved using the CRISPR/Cas9 system. By intercrossing the knockout and NOD mice, we obtained a novel strain of NOD/SCID/IL2 Rγ-/-（NSG） mice,denoted as cNSG（Chinese NSG） mice. Results In addition to the NOD mutation, cNSG mice exhibited a complete absence of T cells, B cells and NK cells. cNSG mice allowed more efficient engraftment of human cancer cells than the commonly used immunodeficient nude mice. Conclusion cNSG mice will provide an important xenotransplantation model for biomedical research.

关 键 词：基因工程 CRISPR/Cas9系统 免疫缺陷小鼠 cNSG品系 移植 

分 类 号：R730.3[医药卫生—肿瘤]