Modulation of gene expression by oligomers targeted to chromosomal DNA
View Patent ↗Synthesis of a target transcript of a gene is selectively increased in a mammalian cell by contacting the cell with a polynucleotide oligomer of 12-28 bases complementary to a region within a target promoter of the gene under conditions whereby the oligomer selectively increases synthesis of the target transcript.
1. A method of selectively increasing synthesis of a human progesterone receptor transcript in a mammalian cell comprising the steps of:
(a) contacting the cell with a polynucleotide oligomer of 12-28 bases comprising SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6 under conditions whereby the oligomer selectively increases synthesis of the target transcript; and
(b) detecting resultant selective increased synthesis of the target gene;
wherein the oligomer is double-stranded RNA.
2. The method of claim 1 wherein the oligomer is a double-stranded RNA of 18-25 bases.
3. The method of claim 1 wherein the oligomer comprises a nucleotide having a 2′ chemical modification.
4. The method of claim 1 wherein the oligomer comprises a serum stability-enhancing chemical modification selected from the group consisting of a phosphorothioate internucleotide linkage, a 2′-O-methyl ribonucleotide, a 2′-deoxy-2′-fluoro ribonucleotide, a 2′-deoxy ribonucleotide, a universal base nucleotide, a 5-C-methyl nucleotide, an inverted deoxyabasic residue incorporation, and a locked nucleic acid.
5. The method of claim 1 wherein the cell is a cultured cell in vitro.
6. The method of claim 1 wherein the cell is a in situ in a host.
7. The method of claim 1 wherein the contacting step is free of viral transduction.
8. The method of claim 1 wherein the contacting step is free of viral transduction and the cell is contacted with a composition consisting essentially of the oligomer.
9. The method of claim 1 wherein the contacting step is free of viral transduction, and there is at least a 2-fold resultant increased synthesis of the target transcript.
10. The method of claim 1 wherein the oligomer is a double-stranded RNA of 18-25 bases, a single region of the target promoter is targeted, and there is at least a 2-fold resultant increased synthesis of the target transcript.
11. The method of claim 1 wherein the cell is contacted with a 1-100 nanomolar concentration of the oligomer.
12. A method of decreasing Cox-2 expression in a cell, the method comprising the steps of:
(a) contacting the cell with polynucleotide oligomer of 12-28 bases complementary to a region located between nucleotides −100 to +25 relative to a transcription start site of a human progesterone receptor (hPR) gene under conditions whereby the oligomer selectively increases synthesis of hPR; and
(b) detecting decreased synthesis of the Cox-2; wherein the oligomer is a double-stranded RNA,
wherein the target hPR promoter region is −11 to +8.