Mutant of adeno-associated virus (aav) capsid protein

Abstract

The present invention provides: a mutant of adeno-associated virus (AAV) capsid protein, which contains at least one amino acid substitution in PLA2 domain when compared with the amino acid sequence for wild-type AAV capsid protein; a nucleic acid encoding the mutant; a cell containing the nucleic acid; a method for producing a recombinant AAV particle, comprising a step of culturing the cell to produce the recombinant AAV particle; a recombinant AAV particle containing the mutant; a composition containing the recombinant AAV particle; and a method for transferring a gene into a target cell, comprising a step of bringing the recombinant AAV particle into contact with the target cell.

1 - 11 . (canceled) 
     
     
         12 . A mutant of an adeno-associated virus (AAV) capsid protein, comprising one or more amino acid replacements in a PLA2 domain as compared with the amino acid sequence of a wild-type AAV capsid protein,
 wherein the mutant is a mutant of AAV other than AAV2, and   wherein the one or more amino acid replacements comprise an amino acid replacement at a position corresponding to position 68 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2.   
     
     
         13 . The mutant of an AAV capsid protein according to  claim 12 , wherein the one or more amino acid replacements comprise an amino acid replacement by valine at a position corresponding to position 68 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2. 
     
     
         14 . The mutant of an AAV capsid protein according to  claim 12 , wherein the one or more amino acid replacements further comprise amino acid replacements at positions corresponding to position 3 and position 6 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2. 
     
     
         15 . The mutant of an AAV capsid protein according to  claim 12 , wherein the one or more amino acid replacements comprise:
 (a) an amino acid replacement by threonine at a position corresponding to position 3 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2,   (b) an amino acid replacement by histidine at a position corresponding to position 6 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2, and   (c) an amino acid replacement by valine at a position corresponding to position 68 in the amino acid sequence of wild-type AAV2 VP1 capsid protein shown by SEQ ID NO:2.   
     
     
         16 . A nucleic acid encoding the mutant of an AAV capsid protein according to  claim 12 . 
     
     
         17 . A cell containing the nucleic acid according to  claim 16 . 
     
     
         18 . A method of producing a recombinant AAV particle, the method comprising a step of culturing the cell according to  claim 17  to produce a recombinant AAV particle. 
     
     
         19 . The method of producing a recombinant AAV particle according to  claim 18 , wherein the cell according to  claim 17  further contains a nucleic acid encoding an AAV Rep protein, a nucleic acid encoding an adenovirus-derived element necessary for formation of an AAV particle, and a nucleic acid having a nucleotide sequence of an AAV genomic DNA. 
     
     
         20 . A recombinant AAV particle containing the mutant of an AAV capsid protein according to  claim 12 . 
     
     
         21 . A composition containing the recombinant AAV particle according to  claim 20 . 
     
     
         22 . A method of introducing a gene into a target cell, the method comprising a step of bringing the recombinant AAV particle according to  claim 20  into contact with a target cell.