ADENO-ASSOCIATED VIRUS (AAV) CLADES, SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR
Sequences of novel adeno-associated virus capsids and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. AAV-mediated delivery of therapeutic and immunogenic genes using the vectors of the invention is also provided.
1 . A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp1 capsid protein having a sequence of amino acids 1 to 736 of SEQ ID NO: 123, or a sequence which is at least 95% identical to the full length of amino acids 1 to 736 of SEQ ID NO: 123; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the gene product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.
2 . The method of claim 1 , wherein the sequence of the vp1 protein is at least 97% identical to the full length of amino acids 1 to 736 of SEQ ID NO: 123.
3 . The method of claim 1 , wherein the sequence of the vp1 protein is at least 99% identical to the full-length of amino acids 1 to 736 of SEQ ID NO: 123.
4 . The method of claim 1 , wherein the sequence of the vp1 protein is the full-length of amino acids 1 to 736 of SEQ ID NO: 123.
5 . The method of claim 1 , wherein the gene product is a low density lipoprotein (LDL) receptor, a high density lipoprotein (HDL) receptor, a very low density lipoprotein (VLDL) receptor, Factor VIII, Factor IX, erythropoietin, ornithine transcarbamylase (OTC), vascular endothelial growth factor (VEGF), glucose-6-phosphatase, alpha-1 antitrypsin, or an antibody.
6 . The method of claim 1 , wherein the molecule is a nucleic acid molecule comprising nucleotides 1 to 2208 of SEQ ID NO: 3, or a nucleotide sequence at least 99% identical to nucleotides 1 to 2208 of SEQ ID NO: 3.
7 . The method of claim 1 , wherein the rep gene is from AAV2.
8 . A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp2 capsid protein having a sequence of amino acids 138 to 736 of SEQ ID NO: 123, or a sequence which is at least 95% identical to the full length of amino acids 138 to 736 of SEQ ID NO: 123; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the gene product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.
9 . The method of claim 8 , wherein the sequence of the vp2 protein is at least 97% identical to the full length of amino acids 138 to 736 of SEQ ID NO: 123.
10 . The method of claim 8 , wherein the sequence of the vp2 protein is at least 99% identical to the full-length of amino acids 138 to 736 of SEQ ID NO: 123.
11 . The method of claim 8 , wherein the sequence of the vp2 protein is the full-length of amino acids 138 to 736 of SEQ ID NO: 123.
12 . The method of claim 8 , wherein the gene product is a low density lipoprotein (LDL) receptor, a high density lipoprotein (HDL) receptor, a very low density lipoprotein (VLDL) receptor, Factor VIII, Factor IX, erythropoietin, ornithine transcarbamylase (OTC), vascular endothelial growth factor (VEGF), glucose-6-phosphatase, alpha-1 antitrypsin, or an antibody.
13 . The method of claim 8 , wherein the molecule is a nucleic acid molecule comprising nucleotides 412 to 2208 of SEQ ID NO: 3, or a nucleotide sequence at least 99% identical to nucleotides 412 to 2208 of SEQ ID NO: 3.
14 . The method of claim 8 , wherein the rep gene is from AAV2.
15 . A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: (a) a molecule encoding the AAV vp3 capsid protein having a sequence of amino acids 203 to 736 of SEQ ID NO: 123, or a sequence which is at least 95% identical to the full length of amino acids 203 to 736 of SEQ ID NO: 123; (b) a functional rep gene; (c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the gene product in a host cell; and (d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.
16 . The method of claim 15 , wherein the sequence of the vp3 protein is at least 97% identical to the full length of amino acids 203 to 736 of SEQ ID NO: 123.
17 . The method of claim 15 , wherein the sequence of the vp3 protein is at least 99% identical to the full-length of amino acids 203 to 736 of SEQ ID NO: 123.
18 . The method of claim 15 , wherein the sequence of the vp3 protein is the full-length of amino acids 203 to 736 of SEQ ID NO: 123.
19 . The method of claim 15 , wherein the gene product is a low density lipoprotein (LDL) receptor, a high density lipoprotein (HDL) receptor, a very low density lipoprotein (VLDL) receptor, Factor VIII, Factor IX, erythropoietin, ornithine transcarbamylase (OTC), vascular endothelial growth factor (VEGF), glucose-6-phosphatase, alpha-1 antitrypsin, or an antibody
20 . The method of claim 15 , wherein the molecule is a nucleic acid molecule comprising nucleotides 607 to 2208 of SEQ ID NO: 3, or a nucleotide sequence at least 99% identical to nucleotides 607 to 2208 of SEQ ID NO: 3.
21 . The method of claim 15 , wherein the rep gene is from AAV2.