Enhanced nucleic acid constructs for eukaryotic gene expression
AssigneeDna2.0, Inc.
InventorsJeremy Minshull, Mark Welch, Sridhar Govindrajan, Kate Caves
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Abstract
The present invention provides polynucleotide vectors for high expression of heterologous genes, and methods for constructing such vectors. Some vectors further comprise novel transposons and transposases that further improve expression. Further disclosed are vectors that can be used in a gene transfer system for stably introducing nucleic acids into the DNA of a cell. The gene transfer systems can be used in methods, for example, but not limited to, gene expression, gene therapy, insertional mutagenesis, or gene discovery.
Claims18
A polynucleotide comprising a transposon comprising inverted repeats of a piggyBac-like transposon flanking a heterologous polynucleotide, the inverted repeats being flanked by copies of the target site 5′-TTAT-3′ at each end, such that the transposon can be excised leaving a single copy of the 5′-TTAT-3′ target site in place of the transposon in the polynucleotide, wherein the transposon comprises at least 16 contiguous nucleotides from SEQ ID NO:
- 1 to provide a copy of the 5′TTAT-3′ target site and one inverted repeat and at least 16 contiguous nucleotides from SEQ. ID NO:
- 2 to provide the other copy of the 5′-TTAT-3′ target site and the other inverted repeat.
A polynucleotide comprising a transposon comprising inverted repeats of a piggyBac-like transposon flanking a heterologous polynucleotide, the inverted repeats being flanked by copies of the target site 5′-TTAT-3′ at each end, such that the transposon can be excised leaving a single copy of the 5′-TTAT-3′ target site in place of the transposon in the polynucleotide, wherein the transposon comprises two inverted terminal repeat (ITR) sequences, each of which is at least 90% identical to SEQ ID NO:
- 32.
A polynucleotide comprising a transposon comprising inverted repeats of a piggyBac-like transposon flanking a heterologous polynucleotide, the inverted repeats being flanked by copies of the target site 5′-TTAT-3′ at each end, such that the transposon can be excised leaving a single copy of the 5′-TTAT-3′ target site in place of the transposon in the polynucleotide, wherein the transposon comprises a sequence that is at least 90% identical to SEQ ID NO:
- 1 and a sequence that is at least 90% identical to SEQ ID NO:
- 2.
Dependencies are inferred from the claim text.