RAG1/2 knockout pigs with severe combined immunodeficiency

Research output: Contribution to journalArticle

Authors

  • Jiao Huang
  • Xiaogang Guo
  • Nana Fan
  • Jun Song
  • Bentian Zhao
  • Zhen Ouyang
  • Zhaoming Liu
  • Yu Zhao
  • Quanmei Yan
  • Xiaoling Yi
  • Axel Schambach
  • Miguel A Esteban
  • Dongshan Yang
  • Huaqiang Yang
  • Liangxue Lai

Colleges, School and Institutes

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.

Details

Original languageEnglish
Pages (from-to)1496-503
Number of pages8
JournalJournal of Immunology
Volume193
Issue number3
Publication statusPublished - 1 Aug 2014

Keywords

  • Anemia, Aplastic, Animals, DNA-Binding Proteins, Disease Models, Animal, Embryo Transfer, Female, Fibroblasts, Gene Knockout Techniques, Gene Targeting, Homeodomain Proteins, INDEL Mutation, Male, Primary Cell Culture, Recombination, Genetic, Severe Combined Immunodeficiency, Sus scrofa, Swine, Swine, Miniature