SCNT-CRISPR-Cas9 Combo Successfully Produces Genome-edited Dogs

Scientists from Korea were able to produce CRISPR-Cas9-mediated genome-edited dogs using somatic cell nuclear transfer (SCNT) technology with the aim of recovering pathogenic mutations in purebred dogs or to generate inbred animal models to study diseases.

There have been two previous cases in which CRISPR-Cas 9 was used for canine research to produce genome-edited dogs, both did not use SCNT-based canine cloning technology. This particular study was performed to establish a procedure for SCNT-based canine genome editing since SCNT-based canine genome editing can maintain the breed, genotypic background, and phenotype of the donor animals, sans the genome-edited target loci. The DJ-1 gene, a multifunction protein that is expressed in almost all cells and tissues, was identified as the genome editing target.

The scientists constructed a CRISPR-Cas 9 vector that targeted the DJ-1. Vector transfection and antibiotic selection were used to establish genome-edited canine fibroblasts. These were then used for canine SCNT to generate two genome-edited dogs that both had confirmed insertion-deletion mutations at the target locus. No off-targets were found at any potential off-target locus. The DJ-1 expression was found to be either downregulated or completely repressed.

This is the first time that the SCNT technology successfully produced genome-edited dogs through CRISPR-Cas9. The method can be used for further studies to produce pathogenic gene-corrected or disease-modeling dogs.

Learn more about the study in BMC Biotechnology.