Method of obtaining a modified adeno-associated virus capsid

A modified AAV capsid for producing viral vectors based on recombinant adeno-associated virus, said modified AAV capsid being produced by a method comprising:

• a) determining amino acids in capsomere protein of the modified AAV, which are located in the interaction interface region of adjacent pentameric subunits of the capsid;
• b) aligning structurally the modified AAV capsid with a template capsid to determine the pairwise correspondence between each amino acid of the modified AAV from the interaction interface region between adjacent pentameric subunits and the nearest amino acid of the template capsid, as a structural analogue of an original amino acid that is considered a potential substitution,
• wherein the template capsid is understood to mean a structurally similar capsid of a virus selected from the family Parvoviridae, which is other than AAV;
• c) comparing, in a pairwise manner, amino acid residues, determined in step b), of the AAV capsid and the template capsid to identify structural differences between the modified AAV capsid and the template capsid in the interaction interface region of adjacent pentameric subunits of the capsid;
• d) selecting positions of amino acid residues for mutagenesis, wherein the amino acid residues for mutagenesis are located in the interaction interface region of adjacent pentameric subunits of the modified AAV capsid and have a structural difference between the modified AAV capsid and the template capsid in the interaction interface region of adjacent pentameric subunits of the capsid;
• e) selecting an amino acid residue for mutagenesis at a position of the positions selected in step d) using one of the following principles steps:
• (i) substituting the initial amino acid in the capsomere protein of the modified AAV for an amino acid that has a greater volume, other than cysteine and methionine;
• (ii) substituting the initial amino acid in the capsomere protein of the modified AAV for an amino acid that has an increased or decreased number of polar contacts between the capsomeres, other than cysteine and methionine;
• f) introducing one or more amino acid substitutions selected in steps d)-e) in the interaction interface region of adjacent pentameric subunits of the modified AAV capsid;
• g) checking the modified AAV capsids produced in step f) for the presence of one or more improved properties as compared to AAV capsid free of these modifications and selecting modified AAV capsids that have one or more improved properties as compared to AAV capsid free of these modifications, wherein the one or more improved properties are selected from:
• increased efficiency of cell transduction,
• increased production of target protein, and
• increased efficiency of generation due to highly efficient production (assembly) of encapsidated viral vectors based on recombinant adeno-associated virus (rAAV).