Aav virions with decreased immunoreactivity and uses therefor

Abstract

Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2.

1 - 43 . (canceled) 
     
     
         44 . A mutated adeno-associated virus (AAV) capsid protein comprising a substitution of one or more of the amino acids occurring at a position corresponding to a position of the full-length AAV-2 VP2 capsid, wherein the one or more amino acids are selected from the group consisting of amino acid:
 124 substituted with a threonine;   126 substituted with an alanine;   127 substituted with an alanine or a leucine;   128 substituted with an alanine or an aspartic acid;   130 substituted with a threonine or an alanine;   131 substituted with a glutamine or an alanine;   132 substituted with a glutamic acid, an alanine, or an asparagine;   133 substituted with an alanine;   134 substituted with a glutamine, a phenylalanine, or an alanine;   188 substituted with an alanine;   190 substituted with an alanine;   191 substituted with a serine;   193 substituted with an alanine;   245 substituted with an alanine;   246 substituted with an alanine;   247 substituted with an alanine;   248 substituted with an alanine;   315 substituted with an alanine;   317 substituted with an alanine;   318 substituted with an alanine;   320 substituted with an alanine;   322 substituted with an alanine;   329 substituted with an arginine;   331 substituted with an alanine;   332 substituted with an alanine;   334 substituted with an alanine;   335 substituted with an alanine;   347 substituted with a cysteine;   350 substituted with a lysine;   354 substituted with an alanine;   355 substituted with a threonine or an alanine;   356 substituted with an arginine;   357 substituted with a glutamic acid, a glutamine, an alanine, or an asparagine;   358 substituted with a lysine;   359 substituted with an alanine;   360 substituted with a histidine, a lysine, or an alanine;   361 substituted with an alanine;   362 substituted with an alanine;   365 substituted with a phenylalanine or an alanine;   366 substituted with a serine or an alanine;   372 substituted with a glutamine, a lysine, a phenylalanine, or an asparagine;   375 substituted with a proline or an alanine;   377 substituted with an alanine;   390 substituted with an alanine;   391 substituted with an alanine;   392 substituted with a glutamic acid, a valine, an alanine, an asparagine, or an isoleucine;   393 substituted with a glutamine, a lysine, an alanine, or an aspartic acid;   394 substituted with a lysine or an alanine;   395 substituted with an alanine;   396 substituted with an alanine;   407 substituted with an alanine;   411 substituted with a lysine or an alanine;   413 substituted with a lysine or an alanine;   418 substituted with an alanine;   419 substituted with an alanine;   437 substituted with an alanine;   438 substituted with an alanine;   448 substituted with a lysine or an alanine;   450 substituted with a lysine or an alanine;   451 substituted with an alanine or lysine;   452 substituted with an alanine;   494 substituted with a proline;   568 substituted with a lysine or an alanine;   569 substituted with an alanine;   571 substituted with a lysine or an alanine; and   592 substituted with a lysine.   
     
     
         45 . The mutated AAV capsid protein of  claim 44 , wherein the one or more amino acids are selected from the group consisting of amino acid:
 126 substituted with an alanine and 127 substituted with a leucine;   130 substituted with an alanine and 131 substituted with an alanine;   334 substituted with an alanine and 360 substituted with a lysine;   334 substituted with an alanine and 450 substituted with an alanine;   334 substituted with an alanine and 568 substituted with an alanine;   334 substituted with an alanine, 360 substituted with a lysine, 394 substituted with an alanine, and 450 substituted with an alanine;   334 substituted with an alanine, 360 substituted with a lysine, and 394 substituted with an alanine;   334 substituted with an alanine, 360 substituted with a lysine, and 450 substituted with an alanine;   360 substituted with a histidine and 361 substituted with an alanine;   360 substituted with a lysine and 450 substituted with an alanine;   361 substituted with an alanine and 358 substituted with a lysine;   361 substituted with an alanine and 494 substituted with a proline;   361 substituted with an alanine and 592 substituted with a lysine;   411 substituted with an alanine and 413 substituted with an alanine;   450 substituted with an alanine, 568 substituted with an alanine, and 571 substituted with an alanine; and   568 substituted with an alanine and 571 substituted with an alanine.   
     
     
         46 . A polynucleotide encoding the mutated protein of  claim 44 . 
     
     
         47 . A polynucleotide encoding the mutated protein of  claim 45 . 
     
     
         48 . A recombinant AAV virion comprising the mutated protein of  claim 44 . 
     
     
         49 . A recombinant AAV virion comprising the mutated protein of  claim 45 . 
     
     
         50 . A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
 (a) providing a recombinant AAV virion according to  claim 48 ;   (b) delivering said recombinant AAV virion to said cell or tissue.   
     
     
         51 . A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
 (a) providing a recombinant AAV virion according to  claim 49 ;   (b) delivering said recombinant AAV virion to said cell or tissue.