Repressor on IFN-λ promoter and siRNA against GATA1, EVI1, and CRX to alter IFN-λ gene activity
The present invention is directed to the identification of a novel repressor located between ˜1.2 kb to ˜1.6 kb from the translation start site of the IFN-λ1 promoter. The present invention provides a method of using siRNAs against ZEB1 (binds to the repressor region) and BLIMP-1 (binds outside the repressor region) and increases the promoter activity of IFN-λ1 (i.e., increases the production of IFN-λ1 protein). siRNAs against ZEB1 mRNA or BLIMP-1 mRNA increase IFN-λ1 gene activity. There is provided a therapeutic application of siRNAs against ZEB1 and BLIMP-1 mRNAs in treating a mammal (including a human) by increasing the production of IFN-λ1 protein that promotes an anti-viral response as well as treats asthma diseases and colon diseases.
1. A method of increasing IFN-λ1 gene activity in an airway epithelial cell, comprising the steps of:
i) providing an airway epithelial cell in need of increasing IFN-λ1 gene activity, said airway epithelial cell is viral-stimulated; and
ii) exposing said airway epithelial cell to a siRNA oligonucleotide targeted against a transcriptional factor mRNA selected from the group consisting of EVI1 and CRX, thereby increasing IFN-λ1 gene activity in said airway epithelial cell, as indicated by an increase in IFN-λ1 protein expression,
wherein said EVI1 mRNA has a nucleotide sequence set forth in Accession No: NM_001105077 and said CRX mRNA has a nucleotide sequence set forth in Accession No: NM_000554.
2. The method of claim 1 , wherein said siRNA oligonucleotide is at least one siRNA oligonucleotide selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 and SEQ ID NO: 63.
3. The method of claim 1 , wherein said siRNA oligonucleotide is at least one siRNA oligonucleotide selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59.
4. The method of claim 1 , wherein said siRNA oligonucleotide is at least one siRNA oligonucleotide selected from the group consisting of SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 and SEQ ID NO: 63.
5. The method of claim 1 , wherein said increased IFN-λ1 protein expression is measured by an increase in IFN-λ1 protein secretion.
6. The method of claim 1 , wherein said increased IFN-λ1 protein expression is measured by an ELISA.
7. The method of claim 1 , wherein said airway epithelial cell is a human airway epithelial cell.
8. The method of claim 1 , wherein said siRNA oligonucleotide is a modified siRNA oligonucleotide.
9. A method of treating a human subject inflicted with an asthmatic disease, comprising the step of administering a therapeutically effective amount of a siRNA oligonucleotide to said human subject, said siRNA oligonucleotide is targeted against a transcriptional factor mRNA selected from the group consisting of EVI1 and CRX, and induces the production of IFN-λ1 protein having an amino acid sequence set forth in GenBank Accession No. NP_742152, and wherein said EVI1 mRNA has a nucleotide sequence set forth in Accession No: NM_001105077 and said CRX mRNA has a nucleotide sequence set forth in Accession No: NM_000554.
10. The method of claim 9 , wherein said siRNA oligonucleotide is at least one oligonucleotide selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 and SEQ ID NO: 63.
11. The method of claim 9 , wherein said siRNA oligonucleotide is at least one oligonucleotide selected from the group consisting of SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58 and SEQ ID NO: 59.
12. The method of claim 9 , wherein said siRNA oligonucleotide is at least one oligonucleotide selected from the group consisting SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62 and SEQ ID NO: 63.