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Programmed DNA destruction by miniature CRISPR-Cas14 enzymes

Lucas B. HarringtonDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USADavid BursteinDepartment of Earth and Planetary Sciences, University of California, Berkeley, CA 94720, USAJanice S. ChenDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USADavid Páez-EspinoDepartment of Energy, Joint Genome Institute, Walnut Creek, CA 94598, USAEnbo MaDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USAIsaac P. WitteDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USAJoshua C. CofskyDepartment of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USANikos C. KyrpidesDepartment of Energy, Joint Genome Institute, Walnut Creek, CA 94598, USAJillian F. BanfieldDepartment of Earth and Planetary Sciences, University of California, Berkeley, CA 94720, USAJennifer A. DoudnaDepartment of Chemistry, University of California, Berkeley, CA 94720, USA

2018en

ABI

Annotatsiya

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.

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Identifikatorlar

DOI: 10.1126/science.aav4294

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