{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T18:12:28Z","timestamp":1791223948036,"version":"4.1.0"},"reference-count":41,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2025,4,6]],"date-time":"2025-04-06T00:00:00Z","timestamp":1743897600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2019YFA0110700"],"award-info":[{"award-number":["2019YFA0110700"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cell Proliferation"],"published-print":{"date-parts":[[2025,9]]},"abstract":"ABSTRACT<\/jats:title>\n \n Gene\u2010edited (GE) pig\u2010to\u2010human xenotransplantation continues to make breakthroughs, but which kind of gene combination is suitable for organ\u2010specific transplantation remains unclear. In this study, we utilised CRISPR\/Cas9 gene editing technology, PiggyBac transposon system, and serial somatic cell cloning technology to develop GTKO\/CMAHKO\/\u03b24GalNT2KO\/hCD46\/hCD55\/hCD59\/hCD39\/hTBM 8 gene\u2010edited cloned (GEC) donor pigs and performed pig\u2010to\u2010non\u2010human primate (NHP) transplantation to evaluate the effectiveness of these GEC pigs. The 8\u2010GEC pigs were obtained by recloning with a 33\u2010day\u2010old 8\u2010GEC fetus with O blood type, which was generated after cell transfection, screening of cell colonies,\u00a0and somatic cell cloning. Molecular identification at DNA, mRNA, and protein levels confirmed successful 8\u2010gene editing. Three copies of transgenes were identified by droplet digital polymerase chain reaction and whole genome sequencing, which were inserted into the introns of pig RFTN1 and MYO10 genes, as well as the intergenic region between PRLR and LOC110257300 genes of these 8\u2010GEC pigs. The 8\u2010GEC pigs also exhibited the ability of germline transmission when mated with our previously generated 4\u2010GEC male pigs. Moreover, antigen\u2013antibody binding assay and complement\u2010dependent cytotoxicity assay demonstrated that 8\u2010gene editing effectively reduced the immune incompatibility and kidney xenograft from 8\u2010GEC pigs survived for 15 and 17\u2009days in two NHPs, respectively. Postoperatively, the recipient serum antibodies IgA, IgG and IgM, complements C3 and C4, coagulation indicators PT, APTT, TT and FIB, as well as most electrolytes and liver function indicators remained relatively stable. Serum creatinine was normal within 10\u2009days post operation. However, the kidney xenograft developed active antibody\u2010mediated rejection at necropsy, characterised by the deposition of antibodies IgG and IgM, as well as complements C4d, C3c and C5b\u2010C9, infiltration of CD68\n +<\/jats:sup>\n macrophages, and micro\u2010thrombotic embolism of glomerular capillaries, etc. In conclusion, we successfully developed fertile 8\u2010GEC pigs, which effectively alleviated immune rejection and exerted life\u2010supporting kidney function in the recipients.\n <\/jats:p>","DOI":"10.1111\/cpr.70028","type":"journal-article","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T03:01:32Z","timestamp":1743994892000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Production and Functional Verification of 8\u2010Gene (\n GGTA1<\/scp>\n ,\n CMAH<\/scp>\n ,\n \u03b24GalNT2<\/scp>\n ,\n hCD46<\/scp>\n ,\n hCD55<\/scp>\n ,\n hCD59<\/scp>\n ,\n hTBM<\/scp>\n ,\n hCD39<\/scp>\n )\u2010Edited Donor Pigs for Xenotransplantation"],"prefix":"10.1111","volume":"58","author":[{"given":"Jiaoxiang","family":"Wang","sequence":"first","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"Faculty of Animal Science and Technology, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaixiang","family":"Xu","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"College of Veterinary Medicine, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Liu","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital of Kunming Medical University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heng","family":"Zhao","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"College of Veterinary Medicine, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Ameen","family":"Jamal","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gen","family":"Chen","sequence":"additional","affiliation":[{"name":"Guizhou Provincial People's Hospital Guiyang China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoying","family":"Huo","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"College of Veterinary Medicine, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang","family":"Yang","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming 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Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanfei","family":"Huang","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital of Kunming Medical University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongfang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"College of Veterinary Medicine, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianxiong","family":"Guo","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengchong","family":"Wang","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"Faculty of Animal Science and Technology, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Liu","sequence":"additional","affiliation":[{"name":"Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University Kunming China"},{"name":"Yunnan Province Xenotransplantation Research Engineering Center, Yunnan Agricultural University Kunming China"},{"name":"Faculty of Animal Science and Technology, Yunnan Agricultural University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siming","family":"Qu","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital of Kunming Medical University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gang","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pharmacology Innovative Institute of Basic Medical Sciences of Zhejiang University Hangzhou 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University Kunming China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,4,6]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2201422"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-023-02471-9"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1097\/MOT.0000000000001146"},{"issue":"10","key":"e_1_2_9_5_1","first-page":"7","article-title":"Pig\u2010To\u2010Human Xenotransplants Take Another Step Forward","volume":"15","author":"Kuehn B. 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Proliferation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/cpr.70028","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,8]],"date-time":"2025-09-08T02:36:49Z","timestamp":1757299009000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/cpr.70028"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,6]]},"references-count":41,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2025,9]]}},"alternative-id":["10.1111\/cpr.70028"],"URL":"https:\/\/doi.org\/10.1111\/cpr.70028","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.05.19.594889","asserted-by":"object"}]},"ISSN":["0960-7722","1365-2184"],"issn-type":[{"value":"0960-7722","type":"print"},{"value":"1365-2184","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,6]]},"assertion":[{"value":"2024-08-21","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication 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