Generation of genome-edited dogs by somatic cell nuclear transfer

Abstract

Abstract Background Canine cloning technology based on somatic cell nuclear transfer (SCNT) combined with genome-editing tools such as CRISPR-Cas9 can be used to correct pathogenic mutations in purebred dogs or to generate animal models of disease. Results We constructed a CRISPR-Cas9 vector targeting canine DJ-1. Genome-edited canine fibroblasts were established using vector transfection and antibiotic selection. We performed canine SCNT using genome-edited fibroblasts and successfully generated two genome-edited dogs. Both genome-edited dogs had insertion-deletion mutations at the target locus, and DJ-1 expression was either downregulated or completely repressed. Conclusion SCNT successfully produced genome-edited dogs by using the CRISPR-Cas9 system for the first time.

Keywords

• Genome-edited dog
• CRISPR-Cas9
• Somatic nuclear transfer
• DJ-1 knock out