Genome editing in mammalian cells using the CRISPR type I-D nuclease.

Keishi Osakabe, Naoki Wada, Emi Murakami, Naoyuki Miyashita, Yuriko Osakabe

Nucleic acids research 2021 Jun 21

filter terms:

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. © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

Citation

Keishi Osakabe, Naoki Wada, Emi Murakami, Naoyuki Miyashita, Yuriko Osakabe. Genome editing in mammalian cells using the CRISPR type I-D nuclease. Nucleic acids research. 2021 Jun 21;49(11):6347-6363