Isolation, Cloning and Characterization of New Adeno-Associated Virus (AAV) Serotypes

Abstract

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

1 - 113 . (canceled) 
     
     
         114 . An isolated nucleic acid molecule comprising a nucleic acid sequence comprising at least 100 contiguous nucleotides from an AAV-X1 virus. 
     
     
         115 . The isolated nucleic acid molecule of  claim 114 , wherein the nucleic acid sequence encodes an immunogenic portion of a protein selected from the group consisting of an AAV-X1 REP and an AAV-X1 CAP protein. 
     
     
         116 . The isolated nucleic acid molecule of  claim 115 , wherein the AAV-X1 REP protein is selected from the group consisting of AAV-X1 REP40, AAV-X1 REP52, AAV-X1 REP68 and AAV-X1 REP78. 
     
     
         117 . The isolated nucleic acid molecule of  claim 115 , wherein the AAV-X1 CAP protein is selected from the group consisting of AAV-X1 VP1, AAV-X1 VP2 and AAV-X1 VP3. 
     
     
         118 . The isolated nucleic acid molecule of  claim 114 , wherein the nucleic acid sequence encodes a protein comprising at least 50 contiguous amino acids from SEQ ID NO: 21 or SEQ ID NO:49. 
     
     
         119 . The isolated nucleic acid molecule of  claim 114 , wherein the nucleic acid sequence is selected from the group consisting of:
 a) a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence at least 80% identical to SEQ ID NO:21, wherein the polypeptide binds an antibody raised against a protein consisting of SEQ ID NO:21; and,   b) a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence at least 80% identical to SEQ ID NO:49, wherein the polypeptide binds an antibody raised against a protein consisting of SEQ ID NO:49.   
     
     
         120 . The isolated nucleic acid molecule of  claim 114 , wherein the nucleic acid sequence is selected from the group consisting of:
 a) a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO:21, wherein the polypeptide binds an antibody raised against a protein consisting of SEQ ID NO:21; and,   b) a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence at least 95% identical to SEQ ID NO:49, wherein the polypeptide binds an antibody raised against a protein consisting of SEQ ID NO:49.   
     
     
         121 . The isolated nucleic acid molecule of  claim 114 , wherein the nucleic acid encodes a protein comprising SEQ ID NO:21 or SEQ ID NO:49. 
     
     
         122 . The isolated nucleic acid molecule of  claim 114 , wherein the molecule is an AAV vector. 
     
     
         123 . An isolated AAV particle comprising the nucleic acid molecule of  claim 114 . 
     
     
         124 . An isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of:
 a) a nucleic acid sequence at least 95% identical to SEQ ID NO:11;   b) a nucleic acid sequence comprising at least 50 contiguous nucleotides from SEQ ID NO:11;   c) a nucleic acid sequence fully complementary to the sequence of a) or b), wherein the polypeptide encoded by the nucleic acid sequence of a) or b) binds an antibody raised against a protein consisting of SEQ ID NO:11;   d) a nucleic acid sequence at least 95% identical to SEQ ID NO:48;   e) a nucleic acid sequence comprising at least 50 contiguous nucleotides from SEQ ID NO:48; and,   f) a nucleic acid sequence fully complementary to the sequence of d) or e), wherein the polypeptide encoded by the nucleic acid sequence of d) or e) binds an antibody raised against a protein consisting of SEQ ID NO:48.   
     
     
         125 . The isolated nucleic acid molecule of  claim 124 , wherein the nucleic acid sequence comprises SEQ ID NO: 11 or SEQ ID NO:48. 
     
     
         126 . The isolated nucleic acid molecule of  claim 124 , wherein the molecule is an AAV vector. 
     
     
         127 . An isolated AAV particle comprising the nucleic acid molecule of  claim 124 . 
     
     
         128 . An isolated protein comprising an amino acid sequence from an AAV-X1 CAP protein or an AAV-X1 REP protein. 
     
     
         129 . The isolated protein of  claim 128 , wherein the amino acid sequence is selected from the group consisting of:
 a) an amino acid sequence comprising at least 50 contiguous amino acids from SEQ ID NO:21;   b) an amino acid sequence at least 80% identical to SEQ ID NO:21, wherein the isolated protein binds an antibody raised against a protein consisting of SEQ ID NO:21;   c) an amino acid sequence comprising at least 50 contiguous amino acids from SEQ ID NO:49; and,   d) an amino acid sequence at least 80% identical to SEQ ID NO:21, wherein the isolated protein binds an antibody raised against a protein consisting of SEQ ID NO:49.   
     
     
         130 . An isolated antibody that selectively binds the isolated protein of  claim 128 . 
     
     
         131 . An AAV particle comprising the isolated protein of  claim 128 . 
     
     
         132 . Use of the isolated protein of  claim 129  to detect antibodies to AAV-X1, comprising:
 a) obtaining an antibody-containing sample; 
 b) contacting the sample with the isolated protein of  claim 128 ; and, 
 c) determining if an antibody-protein reaction occurred, the presence of such a reaction indicating the presence of antibodies to AAV-X1 virus.