RRC ID 65160
Author Osakabe K, Wada N, Murakami E, Miyashita N, Osakabe Y.
Title Genome editing in mammalian cells using the CRISPR type I-D nuclease.
Journal Nucleic Acids Res
Abstract Adoption of CRISPR-Cas systems, such as CRISPR-Cas9 and CRISPR-Cas12a, has revolutionized genome engineering in recent years; however, application of genome editing with CRISPR type I-the most abundant CRISPR system in bacteria-remains less developed. Type I systems, such as type I-E, and I-F, comprise the CRISPR-associated complex for antiviral defense ('Cascade': Cas5, Cas6, Cas7, Cas8 and the small subunit) and Cas3, which degrades the target DNA; in contrast, for the sub-type CRISPR-Cas type I-D, which lacks a typical Cas3 nuclease in its CRISPR locus, the mechanism of target DNA degradation remains unknown. Here, we found that Cas10d is a functional nuclease in the type I-D system, performing the role played by Cas3 in other CRISPR-Cas type I systems. The type I-D system can be used for targeted mutagenesis of genomic DNA in human cells, directing both bi-directional long-range deletions and short insertions/deletions. Our findings suggest the CRISPR-Cas type I-D system as a unique effector pathway in CRISPR that can be repurposed for genome engineering in eukaryotic cells.
Volume 49(11)
Pages 6347-6363
Published 2021-6-21
DOI 10.1093/nar/gkab348
PII 6291161
PMID 34076237
PMC PMC8216271
MeSH CRISPR-Associated Proteins / chemistry CRISPR-Associated Proteins / genetics CRISPR-Associated Proteins / metabolism* CRISPR-Cas Systems* Endodeoxyribonucleases / chemistry Endodeoxyribonucleases / genetics Endodeoxyribonucleases / metabolism* Gene Editing* HEK293 Cells Humans Mutagenesis Mutation
IF 11.502
Human and Animal Cells 293T(RCB2202)