Eukaryotic expression vectors resistant to transgene silencing

摘要

The present invention relates to recombinant DNA molecules such as plasmids, non-viral vectors, viral vectors and hybrids thereof, and more particularly to vector modifications that improve expression of said DNA molecules in cell lines and organisms.

主权项

1. A method of improving the duration of transgene expression, the method comprising: a) providing an expression vector comprising a prokaryotic replication origin, a selectable marker, a eukaryotic expression cassette and tRNA promoters upstream and downstream of the eukaryotic expression cassette, said tRNA promoters each comprising the two intragenic promoter elements RNA pol III type II promoter A-box and B-box, said tRNA promoters preventing gene silencing, said eukaryotic expression cassette comprising the transgene under the control of a heterologous promoter; and b) introducing said expression vector into a target vertebrate cell or a vertebrate organism comprising the target vertebrate cells, under conditions wherein the target vertebrate cells are transfected and said transgene is expressed, wherein the duration of transgene expression is greater than achieved with the same vector lacking said tRNA promoters upstream and downstream of the eukaryotic expression cassette.