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Generation of genome-edited dogs by somatic cell nuclear transfer

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 cani...

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Detalles Bibliográficos
Autores principales: Kim, Dong-Ern, Lee, Ji-Hye, Ji, Kuk-Bin, Park, Kang-Sun, Kil, Tae-Young, Koo, Okjae, Kim, Min-Kyu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9281017/
https://www.ncbi.nlm.nih.gov/pubmed/35831828
http://dx.doi.org/10.1186/s12896-022-00749-3
Descripción
Sumario: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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-022-00749-3.