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Science. 2018 Nov 16;362(6416):839-842. doi: 10.1126/science.aav4294. Epub 2018 Oct 18.

Programmed DNA destruction by miniature CRISPR-Cas14 enzymes.

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
2
Department of Earth and Planetary Sciences, University of California, Berkeley, CA 94720, USA.
3
Department of Energy, Joint Genome Institute, Walnut Creek, CA 94598, USA.
4
Department of Environmental Science, Policy, and Management, University of California, Berkeley, CA 94720, USA.
5
Innovative Genomics Institute, University of California, Berkeley, CA 94720, USA.
6
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA. doudna@berkeley.edu.
7
MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
8
Department of Chemistry, University of California, Berkeley, CA 94720, USA.
9
Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.

Abstract

CRISPR-Cas systems provide microbes with adaptive immunity to infectious nucleic acids and are widely employed as genome editing tools. These tools use RNA-guided Cas proteins whose large size (950 to 1400 amino acids) has been considered essential to their specific DNA- or RNA-targeting activities. Here we present a set of CRISPR-Cas systems from uncultivated archaea that contain Cas14, a family of exceptionally compact RNA-guided nucleases (400 to 700 amino acids). Despite their small size, Cas14 proteins are capable of targeted single-stranded DNA (ssDNA) cleavage without restrictive sequence requirements. Moreover, target recognition by Cas14 triggers nonspecific cutting of ssDNA molecules, an activity that enables high-fidelity single-nucleotide polymorphism genotyping (Cas14-DETECTR). Metagenomic data show that multiple CRISPR-Cas14 systems evolved independently and suggest a potential evolutionary origin of single-effector CRISPR-based adaptive immunity.

PMID:
30337455
DOI:
10.1126/science.aav4294

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