METHODS OF PRODUCING RECOMBINANT VIRAL VECTORS Russian patent published in 2024 - IPC C12N7/02 C12N15/86 

FIELD: biotechnology.

SUBSTANCE: described is a method of producing a recombinant AAV vector. Suitable cell culture is co-transfected with at least two plasmid vectors. First plasmid vector has a plasmid backbone size of more than 5000 base pairs, preferably from 7000 base pairs to 7500 base pairs and contains a heterologous nucleotide sequence flanked by ITR, and an insertion DNA sequence located outside said ITR, preferably adjacent to one ITR, where said insertion sequence has length of 4400 base pairs to 4800 base pairs. Second plasmid vector containing in direction from 5' to 3' coding region rep AAV, coding region cap AAV and nucleotide sequence containing promoter region p5 AAV. Said cells are cultured under conditions enabling AAV replication and packaging. AAV vectors are recovered and the cells are preserved in the cell culture under conditions which ensure further division and growth. Described is a plasmid vector containing: a heterologous nucleotide sequence flanked by ITR, and an intervening DNA sequence located outside said ITR and adjacent to one ITR, where said insertion sequence has length of 4400 base pairs to 4800 base pairs, so that the size of the plasmid backbone is more than 5 thousand base pairs, preferably from 7000 base pairs to 7500 base pairs, wherein said plasmid vector does not contain the F1Ori nucleotide sequence in the backbone sequence, and where said plasmid vector is pcohSgsh-900, as presented in SEQ ID NO: 2. Also described is a plasmid vector containing adenoviral helper functional elements, including sequences VA-RNA, E2A and E4, where said plasmid does not contain the sequence E3, pTB (E2B), ITR Ad and protease, and wherein said plasmid vector is pAdHelper-8615 as presented in SEQ ID NO: 6.

EFFECT: invention extends the range of products for producing recombinant viral vectors.

13 cl, 7 dwg, 1 tbl, 2 ex