title: Efficient generation of B2m-null pigs via injection of zygote with TALENs.
abstract: Donor major histocompatibility complex class I (MHC I) molecules are the main targets of the host immune response after organ allotransplantation. Whether and how MHC I-deficiency of pig donor tissues affects rejection after xenotransplantation has not been assessed. Beta2-microglobulin (B2M) is indispensable for the assembly of MHC I receptors and therefore provides an effective target to disrupt cell surface MHC I expression. Here, we report the one-step generation of mutant pigs with targeted disruptions in B2m by injection of porcine zygotes with B2m exon 2-specific TALENs. After germline transmission of mutant B2m alleles, we obtained F1 pigs with biallelic B2m frameshift mutations. F1 pigs lacked detectable B2M expression in tissues derived from the three germ layers, and their lymphocytes were devoid of MHC I surface receptors. Skin grafts from B2M deficient pigs exhibited remarkably prolonged survival on xenogeneic wounds compared to tissues of non-mutant littermates. Mutant founder pigs with bi-allelic disruption in B2m and B2M deficient F1 offspring did not display visible abnormalities, suggesting that pigs are tolerant to B2M deficiency. In summary, we show the efficient generation of pigs with germline mutations in B2m, and demonstrate a beneficial effect of donor MHC I-deficiency on xenotransplantation.
authors.affiliation: Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China.|School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, Shanghai 201210, China.
authors.forename: Yinan
authors.initials: Y
authors.lastname: Du
authors.affiliation: Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
authors.forename: Lulu
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Immunology, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Institute of Combined Injury, College of Military Preventive Medicine, Third Military Medical University, Chongqing 400038, China.
authors.forename: Fengchao
authors.initials: F
authors.lastname: Wang
authors.affiliation: Research Institute of Burns, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors.forename: Zhengen
authors.initials: Z
authors.lastname: Huang
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China.|School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, Shanghai 201210, China.
authors.forename: Xingxu
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Chongqing 400038, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wei
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods*; D013552:Swine / Q000235:genetics*; D000069896:Transcription Activator-Like Effector Nucleases; D015053:Zygote; D001613:beta 2-Microglobulin / Q000172:deficiency*
chemical_list: D001613:beta 2-Microglobulin; D000069896:Transcription Activator-Like Effector Nucleases