Recombinant aav1, aav5, and aav6 capsid mutants and uses thereof

Abstract

Provided herein are modified recombinant adeno-associated virus (rAAV) capsid proteins, such as modified rAAV1, rAAV5, and rAAV6 capsid proteins, rAAV particles comprising such capsid proteins, nucleic acid molecules encoding such capsid proteins, as well as compositions, kits and methods of use thereof.

What is claimed is: 
     
         1 . A modified adeno-associated virus (AAV) capsid protein, wherein a VP3 region of the modified AAV capsid protein comprises a replacement of tyrosine residues with non-tyrosine residues and/or a replacement of a threonine residue with a non-threonine residue at positions corresponding to:
 Y705, Y731, and T492 of a wild-type AAV1 capsid protein having the sequence of SEQ ID NO: 1,   Y436, Y693, and Y719 of a wild-type AAV5 capsid protein having the sequence of SEQ ID NO: 2, or   Y705, Y731, and T492 of a wild-type AAV6 capsid protein having the sequence of SEQ ID NO: 3.   
     
     
         2 . The modified AAV capsid protein of  claim 1 , wherein the modified AAV capsid protein is a modified AAV1 capsid protein and the modified AAV1 capsid protein comprises replacement of tyrosine residues with non-tyrosine residues and a replacement of a threonine residue with a non-threonine residue at each of the positions corresponding to Y705, Y731, and T492 of the wild-type AAV1 capsid protein having the sequence of SEQ ID NO: 1. 
     
     
         3 . The modified AAV capsid protein of  claim 1 , wherein the modified AAV capsid protein is a modified AAV5 capsid protein and the modified AAV5 capsid protein comprises replacement of tyrosine residues with non-tyrosine residues at each of the positions corresponding to Y436, Y693, and Y719 of a wild-type AAV5 capsid protein having the sequence of SEQ ID NO: 2. 
     
     
         4 . The modified AAV capsid protein of  claim 1 , wherein the modified AAV capsid protein is a modified AAV6 capsid protein and the modified AAV6 capsid protein comprises replacement of tyrosine residues with non-tyrosine residues and a replacement of a threonine residue with a non-threonine residue at each of the positions corresponding to Y705, Y731, and T492 of a wild-type AAV6 capsid protein having the sequence of SEQ ID NO: 3. 
     
     
         5 . The modified capsid protein of any one of  claims 1  to  4 , wherein the non-tyrosine residues are phenylalanine and the non-threonine residue is valine. 
     
     
         6 . A nucleic acid molecule encoding a modified capsid proteins of any one of  claims 1  to  5 . 
     
     
         7 . The nucleic acid molecule of  claim 6 , wherein the nucleic acid molecule is a plasmid. 
     
     
         8 . A recombinant AAV (rAAV) particle comprising a modified capsid protein of any one of  claims 1  to  5 . 
     
     
         9 . A composition comprising a rAAV particle of  claim 8  and a pharmaceutically-acceptable carrier. 
     
     
         10 . A method, comprising:
 contacting a host cell with an rAAV particle comprising the modified AAV1 capsid protein of  claim 2  or  claim 5  or a composition comprising the rAAV particle and a pharmaceutically-acceptable carrier.   
     
     
         11 . The method of  claim 10 , wherein the host cell is a muscle cell. 
     
     
         12 . A method, comprising:
 contacting a host cell with an rAAV particle comprising the modified AAV5 capsid protein of  claim 3  or  claim 5  or a composition comprising the rAAV particle and a pharmaceutically-acceptable carrier.   
     
     
         13 . The method of  claim 12 , wherein the host cell is a retinal or airway epithelial cell. 
     
     
         14 . A method, comprising:
 contacting a host cell with an rAAV particle comprising the modified AAV6 capsid protein of  claim 4  or  claim 5  or a composition comprising the rAAV particle and a pharmaceutically-acceptable carrier.   
     
     
         15 . The method of  claim 14 , wherein the host cell is a hematopoietic stem cell, a dendritic cell, a monocyte, an airway epithelial cell, a muscle cell or a microglial cell. 
     
     
         16 . A kit comprising any of the modified capsid proteins of  claims 1  to  5 , or the rAAV particle of  claim 8 , or the composition of  claim 9 .