DNA vectors and elements for sustained gene expression in eukaryotic cells
The present invention provides polynucleotide vectors for high expression of heterologous genes. Some vectors further comprise novel elements that further improve expression. The gene transfer systems can be used in methods, for example, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery.
1 . A polynucleotide comprising a posttranscriptional regulatory element (PRE) comprising the sequence of SEQ ID NO:313 in operable linkage to an open reading frame or an RNA-encoding sequence, said PRE is positioned 3′ to the open reading frame or RNA-encoding sequence.
2 . The polynucleotide of claim 1 , wherein the polynucleotide further comprises SEQ ID NO: 303, 305 or 307.
3 . The polynucleotide of claim 1 , wherein the sequence of SEQ ID NO:313 is within a sequence comprising SEQ ID NO:290, 291 or 292.
4 . The polynucleotide of claim 1 further comprising a heterologous promoter.
5 . The polynucleotide of claim 1 , further comprising a second open reading frame or second RNA-encoding sequence in operable linkage to a PRE element different than the sequence of SEQ ID NO:313.
6 . A method for expressing an open reading frame or an RNA-encoding sequence comprising introducing into a mammalian cell the polynucleotide of claim 1 .
7 . A mammalian cell comprising the polynucleotide of claim 1 .
8 . A vector comprising the polynucleotide of claim 1 .