Herpesviral lytic gene functions render the viral genome susceptible to novel editing by CRISPR/Cas9

Abstract
Data availability
Article and author information
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Abstract

Herpes simplex virus (HSV) establishes lifelong latent infection and can cause serious human disease, but current antiviral therapies target lytic but not latent infection. We screened for sgRNAs that cleave HSV-1 DNA sequences efficiently in vitro and used these sgRNAs to observe the first editing of quiescent HSV-1 DNA. The sgRNAs targeted lytic replicating viral DNA genomes more efficiently than quiescent genomes, consistent with the open structure of lytic chromatin. Editing of latent genomes caused short indels while editing of replicating genomes produced indels, linear molecules and large genomic sequence loss around the gRNA target site. The HSV ICP0 protein and viral DNA replication increased the loss of DNA sequences around the gRNA target site. We conclude that HSV, by promoting open chromatin needed for viral gene expression and by inhibiting the DNA damage response, makes the genome vulnerable to a novel form of editing by CRISPR-Cas9 during lytic replication.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

The following previously published data sets were used

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Article and author information

Author details

Hyung Suk Oh

Department of Microbiology, Harvard Medical School, Boston, United States

Competing interests
Hyung Suk Oh, We have patent applications pending. U.S. Patent application No. 62/365,826, International Patent Application PCT/US2017/043225.

"This ORCID iD identifies the author of this article:" 0000-0002-1739-0389
Werner M Neuhausser

Department of Obstetrics and Gynecology, Harvard Medical School, Boston, United States

For correspondence
wneuhaus@bidmc.harvard.edu

Competing interests
Werner M Neuhausser, We have patent applications pending. U.S. Patent application No. 62/365,826, International Patent Application PCT/US2017/043225.

"This ORCID iD identifies the author of this article:" 0000-0002-5092-2658
Pierce Eggan

Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States

Competing interests
No competing interests declared.
Magdalena Angelova

Department of Microbiology, Harvard Medical School, Boston, United States

Competing interests
Magdalena Angelova, We have patent applications pending. U.S. Patent application No. 62/365,826, International Patent Application PCT/US2017/043225.
Rory Kirchner

Department of Biostatistics, Harvard T H Chan School of Public Health, Boston, United States

Competing interests
No competing interests declared.
Kevin C Eggan

Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, United States

Competing interests
Kevin C Eggan, We have patent applications pending. U.S. Patent application No. 62/365,826, International Patent Application PCT/US2017/043225.
David M Knipe

Department of Microbiology, Harvard Medical School, Boston, United States

For correspondence
david_knipe@hms.harvard.edu

Competing interests
David M Knipe, Reviewing editor, eLifeWe have patent applications pending. U.S. Patent application No. 62/365,826, International Patent Application PCT/US2017/043225.

"This ORCID iD identifies the author of this article:" 0000-0003-1554-6236

Funding

National Institutes of Health (P01 AI098681)

David M Knipe

National Institutes of Health (R21 AI135423)

Kevin C Eggan

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.