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Title: Programmed DNA destruction by miniature CRISPR-Cas14 enzymes

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.
Authors:
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Publication Date:
Grant/Contract Number:
AC02-05CH11231; AC02–05CH11231
Type:
Published Article
Journal Name:
Science
Additional Journal Information:
Journal Name: Science Journal Volume: 362 Journal Issue: 6416; Journal ID: ISSN 0036-8075
Publisher:
American Association for the Advancement of Science (AAAS)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
OSTI Identifier:
1482289

Harrington, Lucas B., Burstein, David, Chen, Janice S., Paez-Espino, David, Ma, Enbo, Witte, Isaac P., Cofsky, Joshua C., Kyrpides, Nikos C., Banfield, Jillian F., and Doudna, Jennifer A.. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. United States: N. p., Web. doi:10.1126/science.aav4294.
Harrington, Lucas B., Burstein, David, Chen, Janice S., Paez-Espino, David, Ma, Enbo, Witte, Isaac P., Cofsky, Joshua C., Kyrpides, Nikos C., Banfield, Jillian F., & Doudna, Jennifer A.. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes. United States. doi:10.1126/science.aav4294.
Harrington, Lucas B., Burstein, David, Chen, Janice S., Paez-Espino, David, Ma, Enbo, Witte, Isaac P., Cofsky, Joshua C., Kyrpides, Nikos C., Banfield, Jillian F., and Doudna, Jennifer A.. 2018. "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes". United States. doi:10.1126/science.aav4294.
@article{osti_1482289,
title = {Programmed DNA destruction by miniature CRISPR-Cas14 enzymes},
author = {Harrington, Lucas B. and Burstein, David and Chen, Janice S. and Paez-Espino, David and Ma, Enbo and Witte, Isaac P. and Cofsky, Joshua C. and Kyrpides, Nikos C. and Banfield, Jillian F. and Doudna, Jennifer A.},
abstractNote = {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.},
doi = {10.1126/science.aav4294},
journal = {Science},
number = 6416,
volume = 362,
place = {United States},
year = {2018},
month = {10}
}

Works referenced in this record:

Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
journal, October 2015
  • Zetsche, Bernd; Gootenberg, Jonathan�S.; Abudayyeh, Omar�O.
  • Cell, Vol. 163, Issue 3, p. 759-771
  • DOI: 10.1016/j.cell.2015.09.038

An updated evolutionary classification of CRISPR�Cas systems
journal, September 2015
  • Makarova, Kira S.; Wolf, Yuri I.; Alkhnbashi, Omer S.
  • Nature Reviews Microbiology, Vol. 13, Issue 11, p. 722-736
  • DOI: 10.1038/nrmicro3569

IMG/M 4 version of the integrated metagenome comparative analysis system
journal, October 2013
  • Markowitz, Victor M.; Chen, I-Min A.; Chu, Ken
  • Nucleic Acids Research, Vol. 42, Issue D1, p. D568-D573
  • DOI: 10.1093/nar/gkt919

Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems
journal, November 2015

WebLogo: A Sequence Logo Generator
journal, May 2004
  • Crooks, Gavin E.; Hon, Gary; Chandonia, John-Marc
  • Genome Research, Vol. 14, Issue 6, p. 1188-1190
  • DOI: 10.1101/gr.849004