COMPOSITIONS AND METHODS TO TREAT LATENT VIRAL INFECTIONS
Viral infection is a persistent cause of human disease. Guided nuclease systems target the genomes of viral infections, rendering the viruses incapacitated.
1 . A method for treating a viral infection, the method comprising:
introducing into a cell a nuclease and a sequence-specific targeting moiety;
targeting the nuclease to a viral nucleic acid by means of the sequence-specific targeting moiety; and
cleaving the viral nucleic acid with the nuclease without interfering with a host genome.
2 . The method of claim 1 , wherein the nuclease is one selected from the group consisting of a zinc-finger nuclease, a transcription activator-like effector nuclease, and a meganuclease.
3 . The method of claim 1 , wherein the nuclease comprises Cas9 endonuclease and the sequence-specific targeting moiety comprises a guide RNA.
4 . The method of claim 1 , wherein the viral infection is latent and the viral nucleic acid is integrated into the host genome.
5 . The method of claim 1 , wherein cleaving includes making a double strand break in the viral nucleic acid.
6 . The method of claim 1 , wherein the host is a living human subject and the steps are performed in vivo.
7 . The method of claim 1 , further comprising inserting a polynucleotide into the viral nucleic acid.
8 . The method of claim 1 , wherein the viral nucleic acid is from a virus that is selected from the group consisting of Adenovirus, Herpes simplex, type 1, Herpes simplex, type 2, Varicella-zoster virus, Epstein-barr virus, Human cytomegalovirus, Human herpesvirus, type 8, Human papillomavirus, BK virus, JC virus, Smallpox, Hepatitis B virus, Human bocavirus, Parvovirus B19, Human astrovirus, Norwalk virus, coxsackievirus, hepatitis A virus, poliovirus, rhinovirus, Severe acute respiratory syndrome virus, Hepatitis C virus, yellow fever virus, dengue virus, West Nile virus, Rubella virus, Hepatitis E virus, Human immunodeficiency virus (HIV), Influenza virus, Guanarito virus, Junin virus, Lassa virus, Machupo virus, Sabiá virus, Crimean-Congo hemorrhagic fever virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Parainfluenza virus, Respiratory syncytial virus, Human metapneumovirus, Hendra virus, Nipah virus, Rabies virus, Hepatitis D, Rotavirus, Orbivirus, Coltivirus, and Banna virus.
9 . The method of claim 1 , wherein introducing the nuclease and sequence-specific targeting moiety comprises introducing into the cell a viral vector that encodes the nuclease and sequence-specific targeting moiety.
10 . The method of claim 9 , wherein the viral vector is selected from the group consisting of retrovirus, lentivirus, adenovirus, herpesvirus, poxvirus, alphavirus, vaccinia virus and adeno-associated viruses.
11 . The method of claim 1 , wherein introducing the nuclease and sequence-specific targeting moiety comprises introducing into the cell a non-viral vector.
12 . The method of claim 11 , wherein the non-viral vector is selected from the group consisting of a nanoparticle, a cationic lipid, a cationic polymer, metallic nanoparticle, a nanorod, a liposome, microbubbles, a cell-penetrating peptide, and a liposphere.
13 . The method of claim 11 , wherein the non-viral vector comprises polyethyleneglycol (PEG).
14 . The method of claim 11 , further comprising applying energy to the vector.
15 . The method of claim 11 , wherein the energy is introduced via ultrasound or electroporation.
16 . A composition for treatment of a viral infection, the composition comprising:
a nuclease; and
a sequence-specific targeting moiety that targets the nuclease to a viral nucleic acid in vivo within a host cell thereby causing the nuclease to cleave the viral nucleic acid without interfering with host nucleic acid.
17 . The composition of claim 16 , wherein the nuclease is a Cas9 endonuclease and the sequence-specific binding module comprises a guide RNA that specifically targets a portion of a viral genome.
18 . The composition of claim 17 , wherein the Cas9 endonuclease and the guide RNA are co-expressed in a host cell infected by a virus.
19 . The composition of claim 16 , wherein the nuclease is one selected from the list consisting of a zinc-finger nuclease, a transcription activator-like effector nuclease, and a meganuclease.
20 . The composition of claim 16 , wherein the viral nucleic acid is from a virus that is selected from the group consisting of herpes simplex virus (HSV)-1, HSV-2, varicella zoster virus (VZV), cytomegalovirus (CMV), human herpesvirus (HHV)-6, HHV-7, Kaposi's sarcoma-associated herpesvirus (KSHV), JC virus, BK virus, parvovirus b19, adeno-associated virus (AAV), and adenovirus.
21 . A composition for treatment of a viral infection, the composition comprising:
nucleic acid encoding:
a nuclease, and
a sequence-specific targeting moiety that targets the nuclease to viral nucleic acid thereby causing the nuclease to cleave the viral nucleic acid without interfering with host nucleic acid.
22 . The composition of claim 21 , wherein the sequence-specific targeting moiety comprises a guide RNA.
23 . The composition of claim 22 , wherein the guide RNA is complementary to a portion of a viral genome.
24 . The composition of claim 23 , wherein the guide RNA is designed to cause the nuclease to cleave the viral genome within a feature that is necessary for viral function.
25 . The composition of claim 24 , wherein the feature is one selected from the list consisting of: a viral replication origin, a terminal repeat, a replication factor binding site, a promoter, a coding sequence, and a repetitive region.
26 . The composition of claim 21 , wherein the nucleic acid is provided within a delivery vector.
27 . The composition of claim 26 , wherein the delivery vector comprises an adeno-associated virus.
28 . The composition of claim 27 , wherein the delivery vector comprises on selected from the group consisting of retrovirus, lentivirus, adenovirus, herpesvirus, poxvirus, alphavirus, vaccinia virus, a nanoparticle, a cationic lipid, a cationic polymer, a metallic nanoparticle, a nanorod, a liposome, microbubbles, cell-penetrating peptide, a liposphere, and polyethyleneglycol (PEG).
29 . The composition of claim 21 , wherein the nuclease is a Cas9 endonuclease.