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.

What is claimed is:  
     
         1 . A nucleic acid vector comprising a pair of adeno-associated virus 4 (AAV4) inverted terminal repeats and a promoter between the inverted terminal repeats.  
     
     
         2 . A nucleic acid vector comprising a pair of adeno-associated virus 4 (AAV4) inverted terminal repeats and a promoter between the inverted terminal repeats, wherein the AAV4 inverted terminal repeats comprise the nucleotide sequence set forth in SEQ ID NO:6.  
     
     
         3 . A nucleic acid vector comprising a pair of adeno-associated virus 4 (AAV4) inverted terminal repeats and a promoter between the inverted terminal repeats, wherein the AAV4 inverted terminal repeats comprise the nucleotide sequence set forth in SEQ ID NO:20.  
     
     
         4 . The vector of  claim 2 , wherein the promoter is an AAV promoter p5.  
     
     
         5 . The vector of  claim 4 , wherein the p5 promoter is AAV4 p5 promoter.  
     
     
         6 . The vector of  claim 2 , further comprising an exogenous nucleic acid functionally linked to the promoter.  
     
     
         7 . The vector of  claim 2  encapsidated in an adeno-associated virus particle.  
     
     
         8 . The particle of  claim 7 , wherein the particle is an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4.  
     
     
         9 . The particle of  claim 7 , wherein the particle is an AAV1 particle, an AAV2 particle, an AAV3 particle or an AAV5 particle.  
     
     
         10 . An AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4.  
     
     
         11 . The particle of  claim 10 , wherein the vector further comprises an exogenous nucleic acid inserted between the inverted terminal repeats.  
     
     
         12 . An isolated nucleic acid comprising the nucleotide sequence set forth in SEQ ID NO:1.  
     
     
         13 . An isolated nucleic acid consisting essentially of the nucleotide sequence set forth in SEQ ID NO:1.  
     
     
         14 . An isolated nucleic acid that selectively hybridizes with the nucleic acid of  claim 13 .  
     
     
         15 . An isolated nucleic acid encoding an adeno-associated virus 4 Rep protein.  
     
     
         16 . The nucleic acid of  claim 15 , wherein the adeno-associated virus 4 Rep protein has the amino acid sequence set forth in SEQ D NO:2.  
     
     
         17 . The nucleic acid of  claim 15 , wherein the adeno-associated virus 4 Rep protein has the amino acid sequence set forth in SEQ ID NO:8.  
     
     
         18 . The nucleic acid of  claim 15 , wherein the adeno-associated virus 4 Rep protein has the amino acid sequence set forth in SEQ ID NO:9.  
     
     
         19 . The nucleic acid of  claim 15 , wherein the adeno-associated virus 4 Rep protein has the amino acid sequence set forth in SEQ ID NO:10.  
     
     
         20 . The nucleic acid of  claim 15 , wherein the adeno-associated virus 4 Rep protein has the amino acid sequence set forth in SEQ ID NO:11.  
     
     
         21 . The nucleic acid of  claim 15 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:3.  
     
     
         22 . The nucleic acid of  claim 15 , wherein the nucleic acid consists essentially of the nucleotide sequence set forth in SEQ ID NO:3.  
     
     
         23 . An isolated nucleic acid that selectively hybridizes with the nucleic acid of  claim 22 .  
     
     
         24 . The nucleic acid of  claim 15 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:12.  
     
     
         25 . The nucleic acid of  claim 15 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:13.  
     
     
         26 . The nucleic acid of  claim 15 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:14.  
     
     
         27 . The nucleic acid of  claim 15 , wherein the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:15.  
     
     
         28 . An isolated AAV4 Rep protein having the amino acid sequence set forth in SEQ ID NO:2, or a unique fragment thereof.  
     
     
         29 . An isolated AAV4 Rep protein having the amino acid sequence set forth in SEQ ID NO:8, or a unique fragment thereof.  
     
     
         30 . An isolated AAV4 Rep protein having the amino acid sequence set forth in SEQ ID NO:9, or a unique fragment thereof.  
     
     
         31 . An isolated AAV4 Rep protein having the amino acid sequence set forth in SEQ ID NO:10, or a unique fragment thereof.  
     
     
         32 . An isolated AAV4 Rep protein having the amino acid sequence set forth in SEQ ID NO:11, or a unique fragment thereof.  
     
     
         33 . An isolated antibody that specifically binds the protein of  claim 28 .  
     
     
         34 . An isolated AAV4 capsid protein having the amino acid sequence set forth in SEQ ID NO:4.  
     
     
         35 . An isolated antibody that specifically binds the protein of  claim 34 .  
     
     
         36 . An isolated AAV4 capsid protein having the amino acid sequence set forth in SEQ ID NO:16.  
     
     
         37 . An isolated antibody that specifically binds the protein of  claim 36 .  
     
     
         38 . An isolated AAV4 capsid protein having, the amino acid sequence set forth in SEQ ID NO:18.  
     
     
         39 . An isolated antibody that specifically binds the protein of  claim 38 .  
     
     
         40 . An isolated nucleic acid encoding the adeno-associated virus 4 capsid protein of SEQ ID NO:16.  
     
     
         41 . An isolated nucleic acid encoding the adeno-associated virus 4 capsid protein of SEQ ID NO:4.  
     
     
         42 . The nucleic acid of  claim 41 , wherein the nucleic acid comprises the nucleic acid sequence set forth in SEQ D NO:5.  
     
     
         43 . The nucleic acid of  claim 41 , wherein the nucleic acid consists essentially of the nucleic acid sequence set forth in SEQ ID NO:5.  
     
     
         44 . An isolated nucleic acid that selectively hybridizes with the nucleic acid of  claim 39 .  
     
     
         45 . An isolated nucleic acid that selectively hybridizes with the nucleic acid of SEQ ID NO:4.  
     
     
         46 . An isolated nucleic acid comprising the AAV4 p5 promoter comprising nucleotides 130-291 of SEQ ID NO:1.  
     
     
         47 . A method of screening a cell for infectivity by AAV4 comprising contacting the cell with an AAV4 particle comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4 and detecting the presence of the AAV4 particle in the cells.  
     
     
         48 . A method of screening a cell for infectivity by AAV4 comprising contacting the cell with an AAV4 vector comprising an AAV4 particle comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4 and comprising a known nucleic acid, wherein the presence of the AAV4 vector is detected in the cells by detecting the presence of the known nucleic acid.  
     
     
         49 . A method of determining the suitability of an AAV4 vector for administration to a subject comprising administering to an antibody-containing sample from the subject an antigenic protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:2, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 and residues 438-601of SEQ ID NO:2 and detecting an antibody-antigen reaction in the sample, the presence of a reaction indicating the AAV4 vector to be unsuitable for use in the subject.  
     
     
         50 . A method of determining the presence in a subject of an AAV4-specific antibody comprising administering to an antibody-containing sample from the subject an antigenic protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:2, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 and residues 438-601 SEQ ID NO:4 and detecting an antibody-antigen reaction in the sample, the presence of a reaction-indicating the presence of an AAV4-specific antibody in the subject.  
     
     
         51 . A method of delivering a nucleic acid to a cell comprising administering to the cell an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4, containing a vector comprising the nucleic acid inserted between a pair of AAV inverted terminal repeats, thereby delivering the nucleic acid to the cell.  
     
     
         52 . The method of  claim 52 , wherein the AAV inverted terminal repeats are AAV4 inverted terminal repeats.  
     
     
         53 . The method of  claim 52 , wherein the AAV inverted terminal repeats are AAV2 inverted terminal repeats.  
     
     
         54 . A method of delivering a nucleic acid to a subject comprising administering to a cell from the subject an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4, comprising the nucleic acid inserted between a pair of AAV inverted terminal repeats, and returning the cell to the subject, thereby delivering the nucleic acid to the subject.  
     
     
         55 . A method of delivering a nucleic acid to a cell in a subject comprising administering to the subject an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4, comprising the nucleic acid inserted between a pair of AAV inverted terminal repeats, thereby delivering the nucleic acid to a cell in the subject.  
     
     
         56 . A method of delivering a nucleic acid to a cell in a subject having antibodies to AAV2 comprising administering to the subject an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 438-601 shown in SEQ ID NO:4, comprising the nucleic acid, thereby delivering the nucleic acid to a cell in the subject.  
     
     
         57 . The vector of  claim 3 , wherein the promoter is an AAV promoter p5.  
     
     
         58 . The vector of  claim 3 , wherein the p5 promoter is AAV4 p5 promoter.  
     
     
         59 . The vector of  claim 3 , further comprising an exogenous nucleic acid functionally linked to the promoter.  
     
     
         60 . The vector of  claim 3 , encapsidated in an adeno-associated virus particle.  
     
     
         61 . The particle of  claim 61 , wherein the particle is an AAV4 particle, comprising a capsid protein comprising an amino acid sequence defined by amino acids 43 8-601 shown in SEQ ID NO:4.  
     
     
         62 . The particle of  claim 61 , wherein the particle is an AAV1 particle, an AAV2 particle, an AAV3 particle or an AAV5 particle.  
     
     
         63 . An isolated nucleic acid encoding, the adeno-associated virus 4 capsid protein of SEQ ID NO:18.  
     
     
         64 . The particle of  claim 7 , wherein the particle is an AAV4 particle, comprising a capsid protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18.  
     
     
         65 . An AAV4 particle, comprising a capsid protein consisting essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18, containing a vector comprising a pair of AAV2 inverted terminal repeats.  
     
     
         66 . A method of delivering a nucleic acid to a cell comprising administering to the cell an AAV4 particle, comprising a capsid protein consisting essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18, containing a vector comprising the nucleic acid inserted between a pair of AAV-inverted terminal repeats, thereby delivering the nucleic acid to the cell.  
     
     
         67 . A method of delivering a nucleic acid to a subject comprising administering to a cell from the subject an AAV4 particle, comprising a capsid protein consisting essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18, comprising the nucleic acid inserted between a pair of AAV inverted terminal repeats, and returning the cell to the subject, thereby delivering the nucleic acid to the subject.  
     
     
         68 . A method of delivering a nucleic acid to a cell in a subject comprising administering to the subject an AAV4 particle, comprising a capsid protein consisting essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18, comprising the nucleic acid inserted between a pair of AAV inverted terminal repeats, thereby delivering the nucleic acid to a cell in the subject.  
     
     
         69 . A method of delivering a nucleic acid to a cell in a subject having antibodies to AAV2 comprising administering to the subject an AAV4 particle, comprising a capsid protein consisting essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:16 and SEQ ID NO:18, comprising the nucleic acid, thereby delivering the nucleic acid to a cell in the subject.  
     
     
         70 . An AAV4 vector, comprising a nucleic acid selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22.  
     
     
         71 . An isolated adeno-associated virus 4 Rep protein.