High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes

CRISPR/Cas9 has been widely used for the efficient generation of genetically modified animals; however, this system could have unexpected off-target effects. In the present study, we confirmed the validity of a high-fidelity Cas9 variant, HypaCas9, for accurate genome editing in mouse zygotes. HypaC...

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Autores principales: Ikeda, Arisa, Fujii, Wataru, Sugiura, Koji, Naito, Kunihiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6787007/
https://www.ncbi.nlm.nih.gov/pubmed/31633062
http://dx.doi.org/10.1038/s42003-019-0627-8
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author Ikeda, Arisa
Fujii, Wataru
Sugiura, Koji
Naito, Kunihiko
author_facet Ikeda, Arisa
Fujii, Wataru
Sugiura, Koji
Naito, Kunihiko
author_sort Ikeda, Arisa
collection PubMed
description CRISPR/Cas9 has been widely used for the efficient generation of genetically modified animals; however, this system could have unexpected off-target effects. In the present study, we confirmed the validity of a high-fidelity Cas9 variant, HypaCas9, for accurate genome editing in mouse zygotes. HypaCas9 efficiently modified the target locus while minimizing off-target effects even in a single-nucleotide mismatched sequence. Furthermore, by applying HypaCas9 to the discrimination of SNP in hybrid strain-derived zygotes, we accomplished allele-specific gene modifications and successfully generated mice with a monoallelic mutation in an essential gene. These results suggest that the improved accuracy of HypaCas9 facilitates the generation of genetically modified animals.
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spelling pubmed-67870072019-10-18 High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes Ikeda, Arisa Fujii, Wataru Sugiura, Koji Naito, Kunihiko Commun Biol Article CRISPR/Cas9 has been widely used for the efficient generation of genetically modified animals; however, this system could have unexpected off-target effects. In the present study, we confirmed the validity of a high-fidelity Cas9 variant, HypaCas9, for accurate genome editing in mouse zygotes. HypaCas9 efficiently modified the target locus while minimizing off-target effects even in a single-nucleotide mismatched sequence. Furthermore, by applying HypaCas9 to the discrimination of SNP in hybrid strain-derived zygotes, we accomplished allele-specific gene modifications and successfully generated mice with a monoallelic mutation in an essential gene. These results suggest that the improved accuracy of HypaCas9 facilitates the generation of genetically modified animals. Nature Publishing Group UK 2019-10-10 /pmc/articles/PMC6787007/ /pubmed/31633062 http://dx.doi.org/10.1038/s42003-019-0627-8 Text en © The Author(s) 2019 Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Ikeda, Arisa
Fujii, Wataru
Sugiura, Koji
Naito, Kunihiko
High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title_full High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title_fullStr High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title_full_unstemmed High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title_short High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes
title_sort high-fidelity endonuclease variant hypacas9 facilitates accurate allele-specific gene modification in mouse zygotes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6787007/
https://www.ncbi.nlm.nih.gov/pubmed/31633062
http://dx.doi.org/10.1038/s42003-019-0627-8
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