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|Title:||CRISPR-Cas3 induces broad and unidirectional genome editing in human cells|
Center for iPS Cell Research and Application
Institute of Medical Science The University of Tokyo
National Institute of Genetics Mishima
Database Center for Life Science (DBCLS)
|Keywords:||Biochemistry, Genetics and Molecular Biology;Chemistry|
|Citation:||Nature Communications. Vol.10, No.1 (2019)|
|Abstract:||© 2019, The Author(s). 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.|
|Appears in Collections:||Scopus 2019|
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