AVV Vectors and Methods

Abstract

The invention relates to Adeno-associated virus vectors. In particular, it relates to Adeno-associated virus vectors with modified capsid proteins and materials and methods for their preparation and use.

1 . An AAV vector comprising a capsid protein with an amino acid insertion following the capsid amino acid at a position selected from the group consisting of:
 (a) a position corresponding to position 139 in the VP1 capsid of AAV2 (SEQ ID NO: 13)   (b) a position corresponding to position 161 in the VP1 capsid of AAV2 (SEQ ID NO: 13).   (c) a position corresponding to position 459 in the VP1 capsid of AAV2 (SEQ ID NO: 13);   (d) a position corresponding to position 584 in the VP1 capsid of AAV2 (SEQ ID NO: 13);   (e) a position corresponding to position 588 in the VP1 capsid of AAV2 (SEQ ID NO: 13);   (f) a position corresponding to position 657 in the VP1 capsid of AAV2 (SEQ ID NO:   13);   (g) a position corresponding to position 586 in the VP1 capsid of AAV1 (SEQ ID NO: 20);   (h) a position corresponding to position 590 in the VP1 capsid of AAV1 (SEQ ID NO: 20);   (i) a position corresponding to position 586 in the VP1 capsid of AAV3 (SEQ ID NO: 22);   (j) a position corresponding to position 585 in the VP1 capsid of AAV4 (SEQ ID NO: 24); and   (k) a position corresponding to position 575 in the VP1 capsid of AAV5 (SEQ ID NO: 36).   
     
     
         2 . The AAV vector of  claim 1  wherein the AAV vector is selected from the group consisting of AAV1, AAV2, AAV3, AAV4, and AAV5. 
     
     
         3 . The AAV vector of  claim 2  wherein the amino acid insertion comprises a targeting peptide. 
     
     
         4 - 16 . (canceled) 
     
     
         17 . The AAV vector of  claim 2  wherein the amino acid insertion comprises an immunogen. 
     
     
         18 . The AAV vector of  claim 2  wherein the amino acid insertion comprises a substrate for an enzymatic reaction. 
     
     
         19 . The AAV vector of  claim 18  where the substrate is a biotin acceptor peptide. 
     
     
         20 . (canceled) 
     
     
         21 . The AAV vector of  claim 2  wherein the insertion is flanked by a linker/scaffolding sequence. 
     
     
         22 .- 24 . (canceled) 
     
     
         25 . A polynucleotide encoding the capsid protein of an AAV vector of  claim 2 . 
     
     
         26 . A cell transfected with the polynucleotide of  claim 25 . 
     
     
         27 . A method of producing AAV vector comprising a capsid protein with an amino acid insertion, comprising growing a packaging cell and providing the packaging cell with helper virus functions, wherein said packaging cell comprises the polynucleotide of  claim 25 , the AAV rep gene and a recombinant AAV genome comprising DNA of interest flanked by AAV inverted terminal repeats. 
     
     
         28 . The method of  claim 27  wherein said cell expresses biotin ligase. 
     
     
         29 . The method of  claim 28  further comprising the step of treating said AAV vector produced with biotin ligase. 
     
     
         30 . A method of transferring a DNA of interest to a cell comprising delivering to the cell an AAV vector of  claim 2 . 
     
     
         31 . The method of  claim 30  wherein the cell is a cancer cell or an endothelial cell. 
     
     
         32 - 35 . (canceled) 
     
     
         36 . A pharmaceutical composition comprising the AAV vector of  claim 2  in a pharmaceutically acceptable carrier. 
     
     
         37 . An immunogenic composition comprising the AAV vector of  claim 17 . 
     
     
         38 . A method for eliciting an immune response in an animal, said method comprising administering to the animal an immunogenic composition of  claim 37 . 
     
     
         39 . A method of transferring a DNA of interest to a cell comprising delivering an AAV vector encoding the DNA of interest to the cell, wherein said AAV vector comprises a capsid protein containing one or more amino acid insertions that ablate the ability of the vector to bind heparin-sulfate proteoglycan and allow the vector to use a cellular receptor not used by wild type AAV for DNA transfer. 
     
     
         40 . A method of infecting a cell comprising administering an AAV vector to the cell, wherein said AAV vector comprises a capsid protein containing an amino acid insertion, wherein said AAV vector comprises a capsid protein containing one or more amino acid insertions that ablate the ability of the vector to bind heparin-sulfate proteoglycan and allow the vector to use a cellular receptor not used by wild type AAV for infection. 
     
     
         41 . (canceled) 
     
     
         42 . An AAV vector comprising biotinylated capsid protein.