RAG1/2 Knockout Pigs with Severe Combined Immunodeficiency
RAG1/2 Knockout Pigs with Severe Combined Immunodeficiency
复制标题
具有严重联合免疫缺陷的 RAG1/2 基因敲除猪
DOI：
10.4049/jimmunol.1400915
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发表时间：
2014-08-01
影响因子：
4.4
通讯作者：
Lai, Liangxue
中科院分区：
文献类型：
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作者：
Huang, Jiao;Guo, Xiaogang;Lai, Liangxue
猪与人类有许多生理、生化和解剖学上的相似之处，已经成为生物医学研究的有价值的大型动物模型。考虑到在免疫系统相似性，合适的大小，和长寿的临床实践和监测目的的优势，SCID猪携带功能障碍RAG可以作为重要的实验工具，再生医学，同种异体和异种移植物移植，以及重建相关的免疫系统的实验。在这项研究中，我们报告的RAG 1和RAG 2基因敲除猪使用转录激活因子样效应核酸酶的生成和表型特征。利用转录激活因子样效应核酸酶对猪胎儿成纤维细胞进行基因工程改造，然后用于为体细胞核移植提供供体核。我们获得了27只活的克隆仔猪;在这些仔猪中，9只靶向RAG 1中的双等位基因突变，3只靶向RAG 2中的双等位基因突变，10只靶向RAG 2中的单等位基因突变。RAG 1或RAG 2中具有双等位基因突变的仔猪表现出免疫器官发育不全，不能进行V（D）J重排，并且丧失成熟的B和T细胞。这些免疫缺陷RAG 1/2基因敲除猪是生物医学和转化研究的有前途的工具。
Pigs share many physiological, biochemical, and anatomical similarities with humans and have emerged as valuable large animal models for biomedical research. Considering the advantages in immune system resemblance, suitable size, and longevity for clinical practical and monitoring purpose, SCID pigs bearing dysfunctional RAG could serve as important experimental tools for regenerative medicine, allograft and xenograft transplantation, and reconstitution experiments related to the immune system. In this study, we report the generation and phenotypic characterization of RAG1 and RAG2 knockout pigs using transcription activator-like effector nucleases. Porcine fetal fibroblasts were genetically engineered using transcription activator-like effector nucleases and then used to provide donor nuclei for somatic cell nuclear transfer. We obtained 27 live cloned piglets; among these piglets, 9 were targeted with biallelic mutations in RAG1, 3 were targeted with biallelic mutations in RAG2, and 10 were targeted with a monoallelic mutation in RAG2. Piglets with biallelic mutations in either RAG1 or RAG2 exhibited hypoplasia of immune organs, failed to perform V(D)J rearrangement, and lost mature B and T cells. These immunodeficient RAG1/2 knockout pigs are promising tools for biomedical and translational research.