{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T13:03:55Z","timestamp":1779800635654,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,22]]},"group-title":"Bioengineering","reference-count":30,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"NIH","award":["R35GM128645"],"award-info":[{"award-number":["R35GM128645"]}]},{"name":"NIH","award":["R01AI135049"],"award-info":[{"award-number":["R01AI135049"]}]},{"name":"Matthew 6 Fund"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,22]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Alpha-gal Syndrome (AGS) is a potentially life-threatening allergy caused by an IgE-mediated immune response to galactose-\u03b1-1,3-galactose (alpha-gal), a carbohydrate epitope present in most mammalian meats. Currently, strict avoidance of mammalian meat remains the primary management strategy for affected individuals, and alpha-gal-free beef is not commercially available. Here, we leverage cultivated meat as a biotechnology plat-form to address this unmet clinical need by engineering alpha-gal\u2013free bovine muscle cells. Using CRISPR\/Cas9 genome editing, we disrupted GGTA1, the gene encoding \u03b11,3-galactosyltransferase, in immortalized bovine satellite cells (iBSCs). High-efficiency editing produced clonal GGTA1 knockout iBSCs harboring a homozygous frameshift mutation. Flow cytometry and immunofluorescence confirmed loss of the alpha-gal epitope, while bulk RNA-seq indicated minimal disruption of global gene expression and preserved myogenic differentiation capacity. Importantly, lysates from GGTA1 knockout iBSCs elicited substantially reduced basophil activation in assays using plasma from a patient with AGS, indicating reduced basophil activation consistent with reduced allergenic potential. Together, these findings establish a proof of concept for engineering AGS-compatible cultivated meat and demonstrate the potential of cultivated meat technologies to address human health challenges.<\/jats:p>\n                <jats:sec>\n                  <jats:title>HIGHLIGHTS<\/jats:title>\n                  <jats:list list-type=\"simple\">\n                    <jats:list-item>\n                      <jats:label>\u2218<\/jats:label>\n                      <jats:p>CRISPR\/Cas9-mediated disruption of GGTA1 eliminated alpha-gal from bovine satellite cells<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:label>\u2218<\/jats:label>\n                      <jats:p>GGTA1 knockout cells retained myogenic identity and differentiation capacity<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:label>\u2218<\/jats:label>\n                      <jats:p>GGTA1 knockout reduced basophil activation in an alpha-gal syndrome immune assay<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:label>\u2218<\/jats:label>\n                      <jats:p>Genome-edited bovine cells provide a proof of concept for AGS-compatible cultivated meat<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.20.726299","type":"posted-content","created":{"date-parts":[[2026,5,23]],"date-time":"2026-05-23T06:20:21Z","timestamp":1779517221000},"source":"Crossref","is-referenced-by-count":0,"title":["CRISPR-mediated engineering of bovine satellite cells for Alpha-Gal Syndrome-compatible cultivated meat"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8864-2301","authenticated-orcid":false,"given":"Susan","family":"D\u2019Costa","sequence":"first","affiliation":[{"name":"Thurston Arthritis Research Center, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shailesh K.","family":"Choudhary","sequence":"additional","affiliation":[{"name":"Thurston Arthritis Research Center, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Division of Rheumatology, Allergy and Immunology, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grace E.","family":"Kenney","sequence":"additional","affiliation":[{"name":"Curriculum in Bioinformatics and Computational Biology, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacqueline","family":"Shine","sequence":"additional","affiliation":[{"name":"Thurston Arthritis Research Center, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Diekman","sequence":"additional","affiliation":[{"name":"Thurston Arthritis Research Center, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Lampe Joint Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott P.","family":"Commins","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Allergy and Immunology, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Institute for Global Health and Infectious Diseases, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2123-0051","authenticated-orcid":false,"given":"Douglas H.","family":"Phanstiel","sequence":"additional","affiliation":[{"name":"Thurston Arthritis Research Center, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Curriculum in Bioinformatics and Computational Biology, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, NC, United States of America"},{"name":"Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, NC, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052605152559000_2026.05.20.726299v1.1","first-page":"435343","article-title":"Tissue engineering for clean meat production","volume":"3","year":"2019","journal-title":"Frontiers in Sustainable Food 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