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

Author:

Oh Hyung Suk1ORCID,Neuhausser Werner M234ORCID,Eggan Pierce23,Angelova Magdalena1,Kirchner Rory5,Eggan Kevin C236,Knipe David M1ORCID

Affiliation:

1. Department of Microbiology, Blavatnik Institute, Harvard Medical School, Boston, United States

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

3. Harvard Stem Cell Institute, Harvard University, Cambridge, United States

4. Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States

5. Department of Biostatistics, Harvard TH Chan School of Public Health, Boston, United States

6. Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, United States

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.

Funder

National Institutes of Health

Harvard Medical School

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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