Enhanced nucleic acid constructs for eukaryotic gene expression

摘要

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.

主权项

1. A polynucleotide encoding a transposase fused to a heterologous nuclear localization sequence (NLS) and operably linked to a heterologous promoter, wherein the transposase comprises the amino acid sequence of SEQ ID NO: 45 or SEQ ID NO: 46, and wherein the transposase can integrate into a genomic DNA of a host cell at a 5′-TTAA-3′ target sequence a transposon comprising a non-transposon heterologous DNA sequence flanked by a pair of inverted terminal repeats (ITR) of a piggyBac transposon each ITR comprising the SEQ ID NO: 42.