Expression in insect cells of genes with overlapping open reading frames, methods and compositions therefor

摘要

The present teachings disclose nucleic acid cassettes for expressing in an insect cell a plurality of polypeptides encoded by a gene comprising overlapping open reading frames (ORFs). A cassette comprises, in 5′ to 3′ order, a) a first insect cell-operable promoter, b) a 5′ portion of a gene comprising a first ORF of the gene, c) an intron comprising a second insect cell-operable promoter, and d) a 3′ portion of the gene comprising at least one additional ORF. Vectors and insect cells comprising the cassettes are also disclosed, as well as methods for production of recombinant adeno-associated virus in insect cells using the cassettes.

主权项

1. A nucleic acid cassette for expressing in an insect cell a plurality of polypeptides encoded by a parvovirus capsid gene, the cassette comprising in 5′ to 3′ order:
(i) a first insect cell operable promoter linked operably to a 5′ portion of a first ORF of the parvovirus capsid gene, the first ORF comprising a translation initiation codon, (ii) an intron comprising a second insect cell operable promoter, the second promoter operably linked to a 5′ portion of an at least one additional ORF of a parvovirus capsid gene, wherein the at least one additional ORF comprises at least one additional translation initiation codon and overlaps with the 3′ portion of the first ORF.