Aav4 vector and uses thereof

Abstract

The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

1 - 42 . (canceled) 
     
     
         43 . A nucleic acid vector comprising:
 a) a pair of inverted terminal repeats, each of which is capable of forming a T-shaped hairpin structure, and wherein at least one inverted terminal repeat comprises an AAV4 Rep protein binding site and an AAV4 terminal resolution site (trs); and,   b) a heterologous nucleic acid sequence between the terminal repeats.   
     
     
         44 . The nucleic acid vector of  claim 43 , wherein the inverted terminal repeats are AAV4 terminal repeats. 
     
     
         45 . The nucleic acid vector of  claim 43 , wherein the inverted terminal repeats are at least 80% identical in sequence to SEQ ID NO:6 or SEQ ID NO:20, and wherein at least one inverted terminal repeat comprises an AAV4 Rep protein binding site and an AAV4 trs. 
     
     
         46 . The nucleic acid vector of  claim 43 , wherein the inverted terminal repeats are at least 90% identical in sequence to SEQ ID NO:6 or SEQ ID NO:20, and wherein at least one inverted terminal repeat comprises an AAV4 Rep protein binding site and an AAV4 trs. 
     
     
         47 . The nucleic acid vector of  claim 43 , wherein each of the inverted terminal repeats comprises SEQ ID NO:6 or SEQ ID NO:20. 
     
     
         48 . The nucleic acid vector of  claim 43 , wherein the heterologous nucleic acid sequence is functionally linked to a promoter. 
     
     
         49 . The nucleic acid vector of  claim 48 , wherein the promoter is selected from the group consisting of an AAV promoter, an actin gene promoter, an immunoglobulin gene promoter, a cytomegalovirus (CMV) promoter, an adenovirus promoter, a bovine papilloma virus promoter, a heat shock promoter, a respiratory syncytial virus promoter and a Rous Sarcoma virus promoter. 
     
     
         50 . The nucleic acid vector of  claim 48 , wherein the promoter is an AAV2 or an AAV4 promoter. 
     
     
         51 . The nucleic acid vector of  claim 48 , wherein the promoter is an AAV2 p5 promoter or an AAV4 p5 promoter. 
     
     
         52 . The nucleic acid vector of  claim 48 , wherein the promoter comprises a functional portion of SEQ ID NO:7. 
     
     
         53 . The nucleic acid vector of  claim 43 , wherein the heterologous nucleic acid sequence encodes a therapeutic agent. 
     
     
         54 . The nucleic acid vector of  claim 53 , wherein the therapeutic agent is selected from the group consisting of a cytotoxic peptide, a tumor necrosis factors (TNF), an interferon, an interleukin, GM-CSF, adenosine deaminase, a cellular growth factor, soluble CD4, Factor VIII, Factor IX, a T-cell receptors, LDL receptor, ApoE, ApoC, alpha-1 antitrypsin, ornithine transcarbamylase (OTC), cystic fibrosis transmembrane receptor (CFTR), insulin, Fc receptors for antigen binding domains of antibodies, and an antisense RNA. 
     
     
         55 . A recombinant AAV4 particle comprising a nucleic acid vector comprising:
 a) a pair of inverted terminal repeats, each of which is capable of forming a T-shaped hairpin structure, and wherein at least one inverted terminal repeat comprises an AAV4 Rep protein binding site and an AAV4 terminal resolution site (trs); and,   b) a heterologous nucleic acid sequence between the terminal repeats.   
     
     
         56 . The recombinant AAV4 particle of  claim 55 , wherein the inverted terminal repeats are AAV4 terminal repeats. 
     
     
         57 . The recombinant AAV4 particle of  claim 55 , wherein the heterologous nucleic acid sequence is functionally linked to a promoter. 
     
     
         58 . The recombinant AAV4 particle of  claim 57 , wherein the promoter is selected from the group consisting of an AAV promoter, an actin gene promoter, an immunoglobulin gene promoter, a cytomegalovirus (CMV) promoter, an adenovirus promoter, a bovine papilloma virus promoter, a heat shock promoter, a respiratory syncytial virus promoter and a Rous Sarcoma virus (RSV) promoter. 
     
     
         59 . The recombinant AAV4 particle of  claim 55 , wherein the heterologous nucleic acid sequence encodes a therapeutic agent. 
     
     
         60 . The recombinant AAV4 particle of  claim 59 , wherein the therapeutic agent is selected from the group consisting of a cytotoxic peptide, a tumor necrosis factors (TNF), an interferon, an interleukin, GM-CSF, adenosine deaminase, a cellular growth factor, soluble CD4, Factor VIII, Factor IX, a T-cell receptors, LDL receptor, ApoE, ApoC, alpha-1 antitrypsin, ornithine transcarbamylase (OTC), cystic fibrosis transmembrane receptor (CFTR), insulin, Fc receptors for antigen binding domains of antibodies, and an antisense RNA. 
     
     
         61 . A method for treating a subject for a disease, comprising administering to a subject in need of such treatment a recombinant AAV4 particle comprising a nucleic acid vector comprising:
 a) a pair of inverted terminal repeats, each of which is capable of forming a T-shaped hairpin structure, and wherein at least one inverted terminal repeat comprises an AAV4 Rep protein binding site and an AAV4 terminal resolution site (trs); and,   b) a heterologous nucleic acid sequence between the terminal repeats;   wherein the heterologous nucleic acid sequence is functionally linked to a promoter, and wherein the heterologous nucleic acid sequence encodes a therapeutic agent for treating the disease.   
     
     
         62 . The method of  claim 61 , wherein the therapeutic agent is selected from the group consisting of a cytotoxic peptide, a tumor necrosis factors (TNF), an interferon, an interleukin, GM-CSF, adenosine deaminase, a cellular growth factor, soluble CD4, Factor VIII, Factor IX, a T-cell receptors, LDL receptor, ApoE, ApoC, alpha-1 antitrypsin, ornithine transcarbamylase (OTC), cystic fibrosis transmembrane receptor (CFTR), insulin, Fc receptors for antigen binding domains of antibodies, and an antisense RNA.