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Xenotransplantation, especially using pigs as donors, is considered an ideal source of alternative donors due to the high similarity between their organ structures and those of humans. However, significant differences in immune recognition and coagulation regulation between species can easily induce a series of rejection reactions, including hyperacute rejection, acute humoral rejection, T\u2010cell\u2010mediated rejection, and chronic vascular complications. It also carries risks such as physiological metabolic incompatibility and potential viral transmission. In recent years, with the development of tools such as CRISPR\/Cas, precise multi\u2010gene editing technology has become possible, enabling the simultaneous knockout of multiple xenoantigen genes (such as GGTA1, CMAH, and B4GALNT2) and the introduction of human genes regulating complement, coagulation, and immune responses (such as hCD55, hTBM (THBD), and hCD47), significantly improving the immune tolerance and physiological compatibility of donor organs. This article systematically reviews the immune and coagulation barriers in xenotransplantation, focusing on precise multi\u2010gene editing strategies for pigs used in xenotransplantation. It highlights editing pathways such as tandem knock\u2010in at the same site, simultaneous multi\u2010site editing, stepwise modular editing, and homology\u2010directed repair (HDR) enrichment. Combined with representative organ\u2010specific examples (heart, kidney, liver, and lung), including key non\u2010human primate studies and early human exploratory cases where available, it explores the application prospects of these strategies in creating safe clinical\u2010grade donor pigs and promoting the clinical translation of xenotransplantation.<\/jats:p>","DOI":"10.1111\/xen.70128","type":"journal-article","created":{"date-parts":[[2026,4,19]],"date-time":"2026-04-19T22:43:49Z","timestamp":1776638629000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Precise Multi\u2010Gene Editing Strategies for Xenotransplantation Donor Pigs: Overcoming Immune and Coagulation Barriers"],"prefix":"10.1111","volume":"33","author":[{"given":"Qianqian","family":"Li","sequence":"first","affiliation":[{"name":"College of Animal Sciences Zhejiang Key Laboratory of Nutrition and Breeding for High\u2010quality Animal Products Zhejiang University  Hangzhou China"},{"name":"Key Laboratory of Molecular Animal Nutrition (Zhejiang University) Ministry of Education  Hangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-9748-8496","authenticated-orcid":false,"given":"Jingchao","family":"Cao","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory for Animal Genetic Improvement National Engineering Laboratory For Animal Breeding College of Animal Science and Technology Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture China Agricultural University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1187-3037","authenticated-orcid":false,"given":"Shoulong","family":"Deng","sequence":"additional","affiliation":[{"name":"National Science Center for Model Animals China Agricultural University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun","family":"Yu","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory for Animal Genetic Improvement National Engineering Laboratory For Animal Breeding College of Animal Science and Technology Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture China Agricultural University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,19]]},"reference":[{"key":"e_1_2_20_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.1031185"},{"key":"e_1_2_20_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/af\/vfz014"},{"key":"e_1_2_20_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41581\u2010022\u201000624\u20106"},{"key":"e_1_2_20_5_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2120238"},{"key":"e_1_2_20_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010020\u201066311\u20103"},{"key":"e_1_2_20_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/xen.12161"},{"key":"e_1_2_20_8_1","doi-asserted-by":"publisher","DOI":"10.1053\/tmrv.2002.33460"},{"key":"e_1_2_20_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcdb.2017.11.023"},{"key":"e_1_2_20_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20103\u2010642\u201074977\u20103_10"},{"key":"e_1_2_20_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/imr.13147"},{"key":"e_1_2_20_12_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.23.021704.115835"},{"key":"e_1_2_20_13_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.tp.0000226063.36325.02"},{"key":"e_1_2_20_14_1","doi-asserted-by":"publisher","DOI":"10.1172\/jci141374"},{"key":"e_1_2_20_15_1","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600\u20106143.2003.00190.x"},{"key":"e_1_2_20_16_1","first-page":"223","article-title":"Histopathology of Hyperacute Rejection of the Heart: Experimental and Clinical Observations in Allografts and Xenografts","volume":"10","author":"Rose A. 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