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

Abstract

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.

Journal

• Nature communications

  Nature communications 10 5302-, 2019-12-06

  Springer Nature

Keywords

• General Physics and Astronomy
• General Biochemistry, Genetics and Molecular Biology
• General Chemistry
• Multidisciplinary

Details

• CRID: 1050566774754142464
• NII Article ID: 120006848247
• ISSN: 20411723
• DOI: 10.1038/s41467-019-13226-x
• HANDLE: 2433/251038
• Web Site: https://www.nature.com/articles/s41467-019-13226-x.pdf
• Web Site: https://www.nature.com/articles/s41467-019-13226-x
• Text Lang: en
• Article Type: journal article
• Data Source:
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