CRISPR-Mediated Elimination of α-Gal Epitopes in Porcine Myoblasts Reduces Allergenic Potential for Safer Cultivated Meat Production

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Abstract
The cultivated meat industry is attracting growing research interest and significant investment. Red meat allergy affects approximately 3% of the global population, manifesting in complications such as severe itching and hives. While the cultivated meat field has focused on the generation of cell lines and upscaling manufacturing, the allergenicity of cultivated meat remains untested. α-Galactose (α-Gal) is a key allergenic epitope in red meat. Here, we generated GGTA1+/- heterozygous and GGTA1-/- homozygous α-Gal knockout porcine myoblast cell lines using CRISPR-Cas9 to evaluate the allergenicity of cultivated meat, especially pork. We show that long-term culturing of immortalised porcine myoblasts results in accumulation of α-Gal relative to other glycans. Using an in-vitro KU812 basophil activation assay, we further demonstrate that knockout of α-Gal reduces allergenic potential, as evidenced by decreased histamine and leukotriene C4 (LTC4) levels. Our findings provide proof-of-concept that allergenicity can be systematically reduced through targeted genome editing, paving the way for safer cultivated meat products.