title: RAG1/2 knockout pigs with severe combined immunodeficiency.
abstract: 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.
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Jiao
authors: J
authors: Huang
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Xiaogang
authors: X
authors: Guo
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Nana
authors: N
authors: Fan
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Jun
authors: J
authors: Song
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Bentian
authors: B
authors: Zhao
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Zhen
authors: Z
authors: Ouyang
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Zhaoming
authors: Z
authors: Liu
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Yu
authors: Y
authors: Zhao
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Quanmei
authors: Q
authors: Yan
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Xiaoling
authors: X
authors: Yi
authors: Institute of Experimental Hematology, Hannover Medical School, 30625 Hannover, Germany;
authors: Axel
authors: A
authors: Schambach
authors: School of Immunity and Infection, College of Medical and Dental Sciences, University of Birmingham, Birmingham B15 2TT, United Kingdom;
authors: Jon
authors: J
authors: Frampton
authors: Laboratory of Chromatin and Human Disease, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; and Guangdong Stem Cell and Regenerative Medicine Research Centre, Guangzhou Institutes of Biomedicine and Health, Guangzhou 510530, China.
authors: Miguel A
authors: MA
authors: Esteban
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;
authors: Dongshan
authors: D
authors: Yang
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; lai_liangxue@gibh.ac.cn yang_huaqiang@gibh.ac.cn.
authors: Huaqiang
authors: H
authors: Yang
authors: Laboratory of Animal Cloning, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; lai_liangxue@gibh.ac.cn yang_huaqiang@gibh.ac.cn.
authors: Liangxue
authors: L
authors: Lai
mesh_terms: D000741:Anemia, Aplastic / Q000196:embryology / Q000235:genetics / Q000276:immunology; D000818:Animals; D004268:DNA-Binding Proteins / Q000172:deficiency* / Q000235:genetics*; D004195:Disease Models, Animal; D004624:Embryo Transfer; D005260:Female; D005347:Fibroblasts / Q000276:immunology / Q000473:pathology; D055786:Gene Knockout Techniques / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D018398:Homeodomain Proteins / Q000235:genetics*; D054643:INDEL Mutation; D008297:Male; D061251:Primary Cell Culture; D011995:Recombination, Genetic / Q000276:immunology; D016511:Severe Combined Immunodeficiency / Q000196:embryology / Q000235:genetics* / Q000276:immunology*; D034421:Sus scrofa; D013552:Swine; D013556:Swine, Miniature
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein