Mutated internal ribosomal entry site (IRES) for controlled gene expression
AssigneeAgency for Science, Technology and Research
InventorYuansheng Yang
Life of this patent
Estimating the term…
Abstract
The present invention relates to a nucleic acid molecule comprising one or multiple mutant IRES elements. Further, the present invention relates to methods of enhancing gene expression and to methods of differentially controlling expression of one or multiple gene(s) of interest. In addition, the present invention relates to a kit for studying interactions or any application requiring co-expression of multiple genes.
Claims19
Nucleic acid molecule comprising a first internal ribosome entry site (IRES) sequence, wherein the first IRES sequence is a mutant IRES sequence and comprises a nucleotide sequence selected from the group consisting of any one of the nucleotides sequences set forth in SEQ ID Nos. 9-12, 19 and 21.
A method of identifying cells that provide for enhanced gene expression comprising the steps of:
- (i) constructing a cell-based expression system comprising a promoter, a gene encoding for a selection marker, one or more genes of interest different from the selection marker gene, and a mutant IRES sequence that comprises a nucleotide sequence selected from the group consisting of any one of the nucleotides sequences set forth in SEQ ID Nos. 9-12, 19 and 21, wherein the mutant IRES sequence is operably linked to the gene encoding for a selection marker but not the one or more genes of interest;
- (ii) incubating the expression system under conditions that allow expression of the selection marker and the one or more genes of interest;
- and
- (iii) selecting for the selection marker, thus identifying the cells that provide for enhanced gene expression.
Dependencies are inferred from the claim text.