Integration of nucleic acid constructs into eukaryotic cells with a transposase from oryzias
AssigneeDna Twopointo Inc.
InventorsJeremy Minshull, Sridhar Govindarajan, Maggie Lee
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Abstract
The present invention provides polynucleotide vectors for high expression of heterologous genes. 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, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery.
Claims30
A polynucleotide comprising an open reading frame encoding a transposase, the amino acid sequence of which is at least 90% identical to SEQ ID NO:
- 782, operably linked to a heterologous promoter.
A non-naturally occurring polynucleotide encoding a polypeptide, the sequence of which is at least 90% identical to SEQ ID NO:
- 782, wherein the polynucleotide sequence comprises at least 10 synonymous codon differences relative to SEQ ID NO:
- 781 at corresponding positions between the polynucleotide and SEQ ID NO:781.
A method of integrating a transposon into a eukaryotic cell, the method comprising
- (a) introducing into the cell a transposon comprising SEQ ID NO:
- 7 and SEQ ID NO:
- 8 flanking a heterologous polynucleotide at left and right ends respectively;
- and
- (b) introducing into the cell a transposase, the sequence of which is at least 90% identical with SEQ ID NO:
- 782, wherein the transposase transposes the transposon to produce a genome comprising SEQ ID NO:
- 7 and SEQ ID NO:
- 8 flanking the heterologous polynucleotide at left and right ends respectively.
Dependencies are inferred from the claim text.