ZBED6 deficiency enhances porcine carcass traits and meat quality via ISLR-mediated Wnt signaling activation

• ZBED6 deficiency improves growth performance and meat quality of pigs. • ZBED6 deficiency increases the content of n-3PUFA, EAA and FAA in LD muscle. • ISLR is a ZBED6-IGF2-independent and evolutionarily conserved direct target of ZBED6. • ZBED6 deficiency promotes skeletal muscle growth through activates the ISLR/Wnt pathway. Identification of genes that regulate meat production and quality in pigs is crucial for improving the pork industry. We previously created a Zinc finger-BED domain transcription factor (ZBED6)-deficient pig model which exhibited accelerated postnatal growth. Here, we evaluated the effect of ZBED6 on meat quality, flavor, nutritional value, safety and the mechanisms underlying meat production in pigs. Our results indicated that ZBED6 deficiency enlarges body size by enhancing feed efficiency. The results of carcass characteristics and meat quality measurements showed that ZBED6 deficiency enhances carcass lean percentage (46.49±0.62 % for WT vs. 52.70±0.56 % for ZBED6 -/- ; P <0.001) and improves redness (12.39±0.42 for WT vs. 14.53±0.59 for ZBED6 -/- ; P =0.04) and reduces cooking loss (50.34±0.43% for WT vs. 48.34±0.55% for ZBED6 -/- ; P =0.04). Analysis of fatty acid and amino acid profiles showed that ZBED6 deficiency enhances both the nutritional value and flavor of pork. A comprehensive analysis utilizing RNA-seq, quantitative proteomics, and ChIP-seq identified the immunoglobulin superfamily containing leucine-rich repeat (ISLR) as a direct negative target of ZBED6. In C2C12 cells with knockdown of Zbed6 , Islr expression is elevated, activating the canonical Wnt pathway and promoting myoblast differentiation and myotube formation, while knockdown of Islr significantly attenuated these effects. The subchronic oral toxicity study of ZBED6 deficiency pork in rat revealed no significant differences in daily clinical signs, body weight, feed intake, hematology, and serum biochemistry compares to wild-type pork. In summary, our study demonstrates the potential of ZBED6-deficient pigs as a valuable resource for the livestock and food industry, providing new insights into the mechanisms by which ZBED6 promotes muscle growth through the regulation of ISLR pathway.