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CRISPR-Cas3 induces broad and unidirectional genome editing in human cells
Although single-component Class 2 CRISPR systems, such as type II Cas9 or type V Cas12a (Cpf1), are widely used for genome editing in eukaryotic cells, the application of multi-component Class 1 CRISPR has been less developed. Here we demonstrate that type I-E CRISPR mediates distinct DNA cleavage a...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6897959/ https://www.ncbi.nlm.nih.gov/pubmed/31811138 http://dx.doi.org/10.1038/s41467-019-13226-x |
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author | Morisaka, Hiroyuki Yoshimi, Kazuto Okuzaki, Yuya Gee, Peter Kunihiro, Yayoi Sonpho, Ekasit Xu, Huaigeng Sasakawa, Noriko Naito, Yuki Nakada, Shinichiro Yamamoto, Takashi Sano, Shigetoshi Hotta, Akitsu Takeda, Junji Mashimo, Tomoji |
author_facet | Morisaka, Hiroyuki Yoshimi, Kazuto Okuzaki, Yuya Gee, Peter Kunihiro, Yayoi Sonpho, Ekasit Xu, Huaigeng Sasakawa, Noriko Naito, Yuki Nakada, Shinichiro Yamamoto, Takashi Sano, Shigetoshi Hotta, Akitsu Takeda, Junji Mashimo, Tomoji |
author_sort | Morisaka, Hiroyuki |
collection | PubMed |
description | Although single-component Class 2 CRISPR systems, such as type II Cas9 or type V Cas12a (Cpf1), are widely used for genome editing in eukaryotic cells, the application of multi-component Class 1 CRISPR has been less developed. Here we demonstrate that type I-E CRISPR mediates distinct DNA cleavage activity in human cells. Notably, Cas3, which possesses helicase and nuclease activity, predominantly triggered several thousand base pair deletions upstream of the 5′-ARG protospacer adjacent motif (PAM), without prominent off-target activity. This Cas3-mediated directional and broad DNA degradation can be used to introduce functional gene knockouts and knock-ins. As an example of potential therapeutic applications, we show Cas3-mediated exon-skipping of the Duchenne muscular dystrophy (DMD) gene in patient-induced pluripotent stem cells (iPSCs). These findings broaden our understanding of the Class 1 CRISPR system, which may serve as a unique genome editing tool in eukaryotic cells distinct from the Class 2 CRISPR system. |
format | Online Article Text |
id | pubmed-6897959 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-68979592019-12-09 CRISPR-Cas3 induces broad and unidirectional genome editing in human cells Morisaka, Hiroyuki Yoshimi, Kazuto Okuzaki, Yuya Gee, Peter Kunihiro, Yayoi Sonpho, Ekasit Xu, Huaigeng Sasakawa, Noriko Naito, Yuki Nakada, Shinichiro Yamamoto, Takashi Sano, Shigetoshi Hotta, Akitsu Takeda, Junji Mashimo, Tomoji Nat Commun Article Although single-component Class 2 CRISPR systems, such as type II Cas9 or type V Cas12a (Cpf1), are widely used for genome editing in eukaryotic cells, the application of multi-component Class 1 CRISPR has been less developed. Here we demonstrate that type I-E CRISPR mediates distinct DNA cleavage activity in human cells. Notably, Cas3, which possesses helicase and nuclease activity, predominantly triggered several thousand base pair deletions upstream of the 5′-ARG protospacer adjacent motif (PAM), without prominent off-target activity. This Cas3-mediated directional and broad DNA degradation can be used to introduce functional gene knockouts and knock-ins. As an example of potential therapeutic applications, we show Cas3-mediated exon-skipping of the Duchenne muscular dystrophy (DMD) gene in patient-induced pluripotent stem cells (iPSCs). These findings broaden our understanding of the Class 1 CRISPR system, which may serve as a unique genome editing tool in eukaryotic cells distinct from the Class 2 CRISPR system. Nature Publishing Group UK 2019-12-06 /pmc/articles/PMC6897959/ /pubmed/31811138 http://dx.doi.org/10.1038/s41467-019-13226-x 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 Morisaka, Hiroyuki Yoshimi, Kazuto Okuzaki, Yuya Gee, Peter Kunihiro, Yayoi Sonpho, Ekasit Xu, Huaigeng Sasakawa, Noriko Naito, Yuki Nakada, Shinichiro Yamamoto, Takashi Sano, Shigetoshi Hotta, Akitsu Takeda, Junji Mashimo, Tomoji CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title_full | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title_fullStr | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title_full_unstemmed | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title_short | CRISPR-Cas3 induces broad and unidirectional genome editing in human cells |
title_sort | crispr-cas3 induces broad and unidirectional genome editing in human cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6897959/ https://www.ncbi.nlm.nih.gov/pubmed/31811138 http://dx.doi.org/10.1038/s41467-019-13226-x |
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