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Generation of genome-edited dogs by somatic cell nuclear transfer

Dong-Ern Kim¹, Ji-Hye Lee¹, Kuk-Bin Ji¹

¹Laboratory of Animal Reproduction and Physiology, Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, 34134, Korea.

BMC Biotechnology

|July 13, 2022

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:

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

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