책임운영기관 농촌진흥청 국립축산과학원 (로고)

Domestic dogs (Canis lupus familiaris) are the most variable mammalian species. More than 400 breeds have been developed by intense artificial selection from a limited number of founders. Consequently, purebred dogs have a greater risk of suffering from genetic disorders than any other species. A number of scientific publications have described the health problems of purebred dogs and emphasized the need for action; the problem has also been highlighted recently in public media. As a result, many breeders are increasingly using DNA tests to reduce the frequency of deleterious mutations in their breeding programs. However, no direct treatment has been developed to solve these inherent problems. In particular, canine hip dysplasia (HD) is the most common inherited polygenic orthopedic trait in dog; however, there is still no ideal medical or surgical treatment. Genome editing tools, such as CRISPR/Cas9 technology, can be a solution to this genetic problem. In particular, prime editing (PE), a novel and universal precision genome-editing technology has great potential for the correction of pathogenic alleles in purebred dogs. Unlike the conventional CRISPR/Cas9 system, PE does not induce double-strand breaks, which can induce random indel mutations at the target locus. PE was designed to generate a nick in single strands of the target genome locus, and then induce accurate target sequence switching using reverse transcriptase. While PE was originally developed in human cells, it has recently been used to develop genome-edited plant varieties and animals, including mice and fruit flies. However, there is no report on the use of PE in dogs. In this study, we used PE to determine whether point mutations causing canine HD can be corrected in canine fibroblasts. Furthermore, we attempted to confirm the possibility of producing genetically modified dogs by somatic cell nuclear transfer (SCNT) using PE-mediated genetically gene-corrected canine fibroblasts.