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 target transcript of a human major vault protein (hMVP) gene, a E-cadherin gene, a p53 gene, or a PTEN gene in a mammalian cell, the method comprising the steps of:
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; and
detecting resultant selective increased synthesis of the target gene;
wherein the oligomer is double-stranded RNA, and wherein the oligomer is selected from the group consisting of SEQ ID NOS 1-3 and 7-18.
2. The method of claim 1 , wherein the region is located between nucleotides −50 to +25 relative to a transcription start site of the gene.
3. The method of claim 1 , wherein the region is located between nucleotides −30 to +17 relative to a transcription start site of the gene.
4. The method of claim 1 , wherein the region includes a transcription start site of the gene.
5. The method of claim 1 , wherein the target promoter is the promoter of the target transcript.
6. The method of claim 1 , wherein the target promoter is the promoter of an isoform of the target transcript.
7. The method of claim 1 , wherein the target promoter is the promoter of an predetermined isoform of the target transcript, and synthesis of the isoform is inhibited.
8. The method of claim 1 , wherein the target promoter is both the promoter of the target transcript and the promoter of an isoform of the target transcript.
9. The method of claim 1 , wherein the oligomer is double-stranded RNA of 18-25 bases.
10. The method of claim 1 , wherein the oligomer comprises a nucleotide having a 2′ chemical modification.
11. 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.
12. The method of claim 1 , wherein the cell is a cultured cell in vitro.
13. The method of claim 1 , wherein the cell is in situ in a host.
14. The method of claim 1 , wherein the contacting step is free of viral transduction.
15. 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.
16. 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.
17. 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.
18. The method of claim 1 , wherein the cell is contacted with a 1-100 nanomolar concentration of the oligomer.
19. The method of claim 1 , wherein the cell is a cancer cell.