Genome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens

David W. Morgens; Michael Wainberg; Evan A. Boyle; Oana Ursu; Carlos L. Araya; C. Kimberly Tsui; Michael S. Haney; Gaelen T. Hess; Kyuho Han; Edwin E. Jeng; Amy Li; Michael P. Snyder; William J. Greenleaf; Anshul Kundaje; Michael C. Bassik

doi:10.1038/ncomms15178

Nature Communications (May 2017)

Genome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens

David W. Morgens,
Michael Wainberg,
Evan A. Boyle,
Oana Ursu,
Carlos L. Araya,
C. Kimberly Tsui,
Michael S. Haney,
Gaelen T. Hess,
Kyuho Han,
Edwin E. Jeng,
Amy Li,
Michael P. Snyder,
William J. Greenleaf,
Anshul Kundaje,
Michael C. Bassik

Affiliations

David W. Morgens: Department of Genetics, Stanford University
Michael Wainberg: Department of Computer Science, Stanford University
Evan A. Boyle: Department of Genetics, Stanford University
Oana Ursu: Department of Genetics, Stanford University
Carlos L. Araya: Department of Genetics, Stanford University
C. Kimberly Tsui: Department of Genetics, Stanford University
Michael S. Haney: Department of Genetics, Stanford University
Gaelen T. Hess: Department of Genetics, Stanford University
Kyuho Han: Department of Genetics, Stanford University
Edwin E. Jeng: Department of Genetics, Stanford University
Amy Li: Department of Genetics, Stanford University
Michael P. Snyder: Department of Genetics, Stanford University
William J. Greenleaf: Department of Genetics, Stanford University
Anshul Kundaje: Department of Genetics, Stanford University
Michael C. Bassik: Department of Genetics, Stanford University

DOI: https://doi.org/10.1038/ncomms15178
Journal volume & issue: Vol. 8, no. 1
pp. 1 – 8

Abstract

Read online

CRISPR-Cas9 screens are powerful high-throughput tools but can be confounded by nuclease toxicity. Here the authors design a library of variable length gRNAs with thousands of negative controls, including the targeting of ‘safe’ loci to account for on-target site DNA damage toxicity.

Published in Nature Communications

ISSN: 2041-1723 (Online)
Publisher: Nature Portfolio
Country of publisher: United Kingdom
LCC subjects: Science
Website: https://www.nature.com/ncomms/

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