Identification of the CKM Gene as a Potential Muscle-Specific Safe Harbor Locus in Pig Genome

Genetically modified pigs have shown considerable application potential in the fields of life science research and livestock breeding. Nevertheless, a barrier impedes the production of genetically modified pigs. There are too few safe harbor loci for the insertion of foreign genes into the pig genome. Only a few loci ( , and ) have been successfully identified to achieve the ectopic expression of foreign genes and produce gene-edited pigs. Here, we use CRISPR/Cas9-mediated homologous directed repair (HDR) to accurately knock the exogenous gene-of-interest fragments into an endogenous gene in the porcine satellite cells. After porcine satellite cells are induced to differentiate, the gene promoter simultaneously initiates the expression of the gene and the exogenous gene. We infer preliminarily that the gene can be identified as a potential muscle-specific safe harbor locus in pigs for the integration of exogenous gene-of-interest fragments.