CRISPR-mediated engineering of bovine satellite cells for AGS-compatible cultivated meat

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 platform to address this unmet clinical need by engineering alpha-gal–free bovine muscle cells.

Using CRISPR/Cas9 genome editing, we disrupted GGTA1, the gene encoding α1,3-galactosyltransferase, in immortalized bovine satellite cells. High-efficiency editing produced clonal GGTA1 knockout cell lines 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.