[HTML][HTML] Inhibition of HSV-1 replication by gene editing strategy

PC Roehm, M Shekarabi, HS Wollebo, A Bellizzi… - Scientific reports, 2016 - nature.com
PC Roehm, M Shekarabi, HS Wollebo, A Bellizzi, L He, J Salkind, K Khalili
Scientific reports, 2016nature.com
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 …
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.
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