title: Inhibition of HSV-1 Replication by Gene Editing Strategy.
abstract: HSV-1 induced illness affects greater than 85% of adults worldwide with no permanent curative therapy. We used RNA-guided CRISPR/Cas9 gene editing to specifically target for deletion of DNA sequences of the HSV-1 genome that span the region directing expression of ICP0, a key viral protein that stimulates HSV-1 gene expression and replication. We found that CRISPR/Cas9 introduced InDel mutations into exon 2 of the ICP0 gene profoundly reduced HSV-1 infectivity in permissive human cell culture models and protected permissive cells against HSV-1 infection. CRISPR/Cas9 mediated targeting ICP0 prevented HSV-1-induced disintegration of promonocytic leukemia (PML) nuclear bodies, an intracellular event critical to productive HSV-1 infection that is initiated by interaction of the ICP0 N-terminus with PML. Combined treatment of cells with CRISPR targeting ICP0 plus the immediate early viral proteins, ICP4 or ICP27, completely abrogated HSV-1 infection. We conclude that RNA-guided CRISPR/Cas9 can be used to develop a novel, specific and efficacious therapeutic and prophylactic platform for targeted viral genomic ablation to treat HSV-1 diseases.
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.|Department of Otolaryngology/Head and Neck Surgery, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.|Department of Neurosurgery, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Pamela C
authors.initials: PC
authors.lastname: Roehm
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Masoud
authors.initials: M
authors.lastname: Shekarabi
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Hassen S
authors.initials: HS
authors.lastname: Wollebo
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Bellizzi
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Lifan
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Julian
authors.initials: J
authors.lastname: Salkind
authors.affiliation: Department of Neuroscience, Center for Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia, 19140 PA, USA.
authors.forename: Kamel
authors.initials: K
authors.lastname: Khalili
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D002460:Cell Line; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000235:genetics; D000072669:Gene Editing / Q000379:methods*; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D054643:INDEL Mutation; D017384:Sequence Deletion; D014779:Virus Replication
chemical_list: D004279:DNA, Viral