title: Creating class I MHC-null pigs using guide RNA and the Cas9 endonuclease.
abstract: Pigs are emerging as important large animal models for biomedical research, and they may represent a source of organs for xenotransplantation. The MHC is pivotal to the function of the immune system in health and disease, and it is particularly important in infection and transplant rejection. Pigs deficient in class I MHC could serve as important reagents to study viral immunity as well as allograft and xenograft rejection. In this study, we report the creation and characterization of class I MHC knockout pigs using the Cas9 nuclease and guide RNAs. Pig fetal fibroblasts were genetically engineered using Cas9 and guide RNAs, and class I MHC(-) cells were then used as nuclear donors for somatic cell nuclear transfer. We produced three piglets devoid of all cell surface class I proteins. Although these animals have reduced levels of CD4(-)CD8(+) T cells in peripheral blood, the pigs appear healthy and are developing normally. These pigs are a promising reagent for immunological research.
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Luz M
authors.initials: LM
authors.lastname: Reyes
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Jose L
authors.initials: JL
authors.lastname: Estrada
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Zheng Yu
authors.initials: ZY
authors.lastname: Wang
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Blosser
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Rashod F
authors.initials: RF
authors.lastname: Smith
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Richard A
authors.initials: RA
authors.lastname: Sidner
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Leela L
authors.initials: LL
authors.lastname: Paris
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Ross L
authors.initials: RL
authors.lastname: Blankenship
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Caitlin N
authors.initials: CN
authors.lastname: Ray
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202;
authors.forename: Aaron C
authors.initials: AC
authors.lastname: Miner
authors.affiliation: Indiana University Health, Indianapolis, IN 46202; and mtector@iuhealth.org.
authors.forename: Matthew
authors.initials: M
authors.lastname: Tector
authors.affiliation: Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202; Indiana University Health Transplant Institute, Indianapolis, IN 46202.
authors.forename: A Joseph
authors.initials: AJ
authors.lastname: Tector
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004720:Endonucleases / Q000378:metabolism; D005347:Fibroblasts / Q000502:physiology*; D055785:Gene Knockdown Techniques / Q000379:methods*; D005818:Genetic Engineering; D006084:Graft Rejection; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D023421:Models, Animal; D053652:Nuclear Transfer Techniques; D013552:Swine / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D014183:Transplantation, Heterologous; D014777:Virus Diseases / Q000276:immunology*; D000094704:RNA, Guide, CRISPR-Cas Systems
chemical_list: D004720:Endonucleases; D015395:Histocompatibility Antigens Class I; D000094704:RNA, Guide, CRISPR-Cas Systems