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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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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
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author Kim, Dong-Ern
Lee, Ji-Hye
Ji, Kuk-Bin
Park, Kang-Sun
Kil, Tae-Young
Koo, Okjae
Kim, Min-Kyu
author_facet Kim, Dong-Ern
Lee, Ji-Hye
Ji, Kuk-Bin
Park, Kang-Sun
Kil, Tae-Young
Koo, Okjae
Kim, Min-Kyu
author_sort Kim, Dong-Ern
collection PubMed
description 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.
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spelling pubmed-92810172022-07-15 Generation of genome-edited dogs by somatic cell nuclear transfer Kim, Dong-Ern Lee, Ji-Hye Ji, Kuk-Bin Park, Kang-Sun Kil, Tae-Young Koo, Okjae Kim, Min-Kyu BMC Biotechnol Research 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. BioMed Central 2022-07-13 /pmc/articles/PMC9281017/ /pubmed/35831828 http://dx.doi.org/10.1186/s12896-022-00749-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Kim, Dong-Ern
Lee, Ji-Hye
Ji, Kuk-Bin
Park, Kang-Sun
Kil, Tae-Young
Koo, Okjae
Kim, Min-Kyu
Generation of genome-edited dogs by somatic cell nuclear transfer
title Generation of genome-edited dogs by somatic cell nuclear transfer
title_full Generation of genome-edited dogs by somatic cell nuclear transfer
title_fullStr Generation of genome-edited dogs by somatic cell nuclear transfer
title_full_unstemmed Generation of genome-edited dogs by somatic cell nuclear transfer
title_short Generation of genome-edited dogs by somatic cell nuclear transfer
title_sort generation of genome-edited dogs by somatic cell nuclear transfer
topic Research
url 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
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