METHOD OF PRODUCING MODIFIED CAPSID OF ADENO-ASSOCIATED VIRUS Russian patent published in 2024 - IPC C12N15/864 

FIELD: biotechnology.

SUBSTANCE: present application relates to gene therapy and molecular biology. Described is a method of producing modified AAV capsid, which involves: a) determination of amino acids of the capsomer protein of the modified AAV, located in the region of the interface of interaction of adjacent pentameric capsid subunits; b) structural alignment of the modified AAV capsid with the capsid-template to determine the pairwise correspondence between each amino acid of the modified AAV from the region of the interaction interface between neighbouring pentameric subunits and the nearest amino acid of the capsid-template as a structural analogue of the initial amino acid, which is considered to be a potential replacement; c) pairwise comparison of the amino acid residues of the modified AAV and the capsid-template determined at step b) to detect structural differences of the capsid of the modified AAV and the capsid-template in the region of the interface of the neighbouring pentameric subunits of the capsid; d) selection of positions for mutagenesis, wherein amino acid residues for mutagenesis are located in the region of interaction of neighbouring pentameric subunits of the capsid of the modified AAV and have a structural difference between the capsid of the modified AAV and the capsid-template in the interface region of the interaction of adjacent pentameric subunits of the capsid; e) selection of amino acid residue for mutagenesis in position selected at step d); f) introducing one or more amino acid substitutions selected at steps d) - e) in the region of the interface of interaction of neighbouring pentameric subunits of the capsid of the modified AAV. Disclosed is a modified AAV capsid for producing viral vectors based on a recombinant adeno-associated virus, which includes one or several amino acid substitutions in the region of the interaction of neighbouring pentameric subunits of the capsid of the modified AAV. Also disclosed is a vector based on a recombinant adeno-associated virus for delivering a heterologous nucleic acid sequence to a subject, which includes: a modified AAV capsid and a heterologous nucleic acid sequence containing regulatory sequences which provide expression of a product encoded by the heterologous nucleic acid sequence in target cells.

EFFECT: invention extends the range of heterologous nucleic acid delivery means.

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