Engineered viral vector reduces induction of inflammatory and immune responses

Abstract

Modified viral genomes are able to reduce induction of inflammatory and immune anti-viral responses. This manifests itself in reduced NF-kB activity, increased viral transduction rates, and increased expression of transgenes. Viral genomes are modified by incorporating one or more oligonucleotide sequences which are able to bind to TLR9 but not induce activation of it. The oligonucleotide sequences may be synthetic, bacterial, human, or from any other source.

1 . (canceled) 
     
     
         2 . A recombinant viral genome comprising four repeated monomers of TTAGGG (SEQ ID NO: 6). 
     
     
         3 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is an adeno-associated virus (AAV) genome. 
     
     
         4 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is selected from the group consisting of adenovirus, herpes simplex virus, varicella, variola virus, hepatitis B, cytomegalovirus, JC polyomavirus, BK polyomavirus, monkeypox virus, Herpes Zoster, Epstein-Barr virus, human herpes virus 7, Kaposi's sarcoma-associated herpesvirus, and human parvovirus B19. 
     
     
         5 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is single stranded. 
     
     
         6 . The recombinant viral genome of  claim 2 , wherein the viral genome is packaged in a virion. 
     
     
         7 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome comprises a therapeutic gene. 
     
     
         8 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is a cytotoxic virus. 
     
     
         9 - 14 . (canceled) 
     
     
         15 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome comprises a non-human gene. 
     
     
         16 . The recombinant viral genome of  claim 2 , wherein the four repeated monomers of TTAGGG (SEQ ID NO: 6) are inserted downstream of or in a 3′ untranslated region of the viral genome. 
     
     
         17 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is covalently linked by a phosphodiester bond to a nucleic acid sequence comprising the four repeated monomers of TTAGGG (SEQ ID NO: 6). 
     
     
         18 . The recombinant viral genome of  claim 2  further comprising a detectable marker. 
     
     
         19 . The recombinant viral genome of  claim 18 , wherein expression of the detectable marker is inducible. 
     
     
         20 . The recombinant viral genome of  claim 2  comprising the nucleotide sequence of SEQ ID NO: 9. 
     
     
         21 - 42 . (canceled) 
     
     
         43 . The recombinant viral genome of  claim 2  comprising three repeated monomers of the nucleotide sequence of SEQ ID NO: 9. 
     
     
         44 . The recombinant viral genome of  claim 2  comprising five repeated monomers of the nucleotide sequence of SEQ ID NO: 9. 
     
     
         45 . A method of treating a mammal, comprising:
 administering to the mammal the recombinant viral genome of  claim 2 .   
     
     
         46 - 73 . (canceled) 
     
     
         74 . A method of producing a recombinant viral genome, comprising:
 inserting into a viral genome four repeated monomers of TTAGGG (SEQ ID NO: 6) or the nucleotide sequence of SEQ ID NO: 9.   
     
     
         75 . A nucleic acid vector comprising the nucleotide sequence of SEQ ID NO: 9. 
     
     
         76 - 90 . (canceled) 
     
     
         91 . The recombinant viral genome of  claim 2 , wherein the recombinant viral genome is self-complementary. 
     
     
         92 . A recombinant viral genome comprising a sequence that is at least 95% identical to the sequence of SEQ ID NO: 9.