Compositions and Methods of Delivering Treatments for Latent Viral Infections

Abstract

Viral infection is a persistent cause of human disease. Guided nuclease systems of the invention target the genomes of viral infections, rendering the viruses incapacitated. The invention further provides delivery methods and compositions for antiviral therapeutics. Methods and compositions are provided for targeted delivery of antiviral therapeutics into cells of interest using, for example, viral vectors such as adenovirus, AAV, and replication incompetent HSV. These and other delivery systems can be used as vehicles to deliver DNA vectors encoding a nuclease or a cell-killing gene. These delivery methods can also be used to deliver naked DNA or RNA, protein products, plasmids containing a promoter that is active only in a latent viral state which drives a cell-killing gene, or other therapeutic agents.

1 . A composition, wherein said composition is a CRISPR/Cas9/gRNA complex comprising guide RNAs for use in the treatment of latent Herpes viruses within a cell. 
     
     
         2 - 3 . (canceled) 
     
     
         4 . The composition of  claim 1 , wherein the latent Herpes viruses are Epstein-Barr viruses (EBV). 
     
     
         5 . The composition of  claim 1 , wherein the composition causes insertions, deletions, or rearrangements within the viral genome in order to incapacitate or destroy the virus. 
     
     
         6 . The composition of  claim 4 , wherein the CRISPR/Cas9/gRNA complex targets an Epstein-Barr virus (EBV) genomic region selected from the group consisting of EBNA1, EBNA3C, Pst1 repeats, EBNA-LP repeats and 125 bp repeats. 
     
     
         7 . A composition comprising a CRISPR/Cas9 plasmid comprising or consisting of a U6 promoter driven chimeric guide RNA (sgRNA) and a ubiquitous promoter driven Cas9. 
     
     
         8 . The composition of  claim 7 , wherein the composition is transfected into cells by a viral vector or a non-viral vector. 
     
     
         9 . The composition of  claim 7 , wherein the plasmid comprises an EBV origin of replication (EBV oriP). 
     
     
         10 . A composition comprising guide RNAs that target an Epstein-Barr virus (EBV) genomic region selected from the group consisting of EBNA1, EBNA3C, LMP1, PstI repeats, EBNA-LP repeats and 125 bp repeats. 
     
     
         11 . The composition of  claim 10 , wherein the guide RNAs target the following Epstein-Barr virus (EBV) genomic repeat regions: PstI repeats, EBNA-LP repeats and 125 bp repeats.