Method to enhance the transcription regulation of SUPT4H on genes containing repetitive nucleotide sequences
The present invention provides a method of modulating the expression of a gene containing expanded nucleotide repeats in a cell, comprising: inhibiting the biological activity of SPT4 or SUPT4H; and regulating the formation of R-loops. The inhibition step can effectively reduce the expression of the gene containing the expanded nucleotide repeats and the regulatory step can further enhance the inhibition step. The inhibition step and the regulation step are for the purpose of regulating gene expression by interfering the capacity of RNA polymerase II transcribing over a DNA template with lengthy nucleotide repeats.
1. A method of modulating expression of a gene containing expanded tri-nucleotide repeats in a cell, comprising:
administering to the cell a composition comprising an oligonucleotide that is complementary to a nucleic acid encoding SPT4 or SUPT4H, wherein the oligonucleotide is a siRNA, a shRNA, an antisense oligonucleotide, or a chemical reagent that is 6-chloropurine riboside; and
administering a compound that enhances R-loop formation, wherein the compound is selected from a topoisomerase inhibitor or an RNase H inhibitor,
wherein the gene contains a segment of DNA with CAG or CTG tri-nucleotide repeats that are prone to R-loop formation.
2. The method of claim 1 , wherein the oligonucleotide is SUPT4H siRNA.
3. The method of claim 1 , wherein the topoisomerase inhibitor is topotecan.
4. The method of claim 1 , wherein the RNase H inhibitor is tropolone.
5. A method of inhibiting transcription of a gene containing expanded tri-nucleotide repeats in a cell, comprising:
administering to the cell a compound selected from an oligonucleotide that is complementary to a nucleic acid encoding SPT4 or SUPT4H, wherein the oligonucleotide is a siRNA, a shRNA, an antisense oligonucleotide, or a chemical reagent 6-chloropurine riboside; and
administering a compound that enhances R-loop formation, wherein the compound that enhances R-loop formation is selected from a topoisomerase inhibitor or an RNase H inhibitor,
wherein the gene contains a segment of DNA with CAG or CTG tri-nucleotide repeats that are prone to R-loop formation.
6. The method of claim 5 , wherein the oligonucleotide is SUPT4H siRNA.
7. The method of claim 5 , wherein the RNase H inhibitor is tropolone.
8. The method of claim 5 , wherein the topoisomerase inhibitor is topotecan.
9. A method of enhancing drug therapy of a tri-nucleotide repeat expansion disease in a subject in need thereof, comprising administering to said subject an R-loop regulating compound with a tri-nucleotide repeat expansion drug, wherein the tri-nucleotide repeat expansion drug is an oligonucleotide that is complementary to a nucleic acid encoding SPT4 or SUPT4H, wherein the oligonucleotide is a siRNA, a shRNA, an antisense oligonucleotide, or the drug is 6-chloropurine riboside, wherein the R-loop regulating compound is selected from a topoisomerase inhibitor or an RNase H inhibitor, wherein the gene contains a segment of DNA with CAG or CTG tri-nucleotide repeats that are prone to R-loop formation.
10. The method of claim 9 , wherein the tri-nucleotide repeat expansion disease is selected from the group consisting of spinocerebellar ataxia type 1, 2, 3, 7, 17, dentatorubral-pallidoluysian atrophy, spinal bulbar muscular atrophy, myotonic atrophy type 1 and Huntington's disease.
11. The method of claim 9 , wherein the oligonucleotide is SUPT4H siRNA, the RNase H inhibitor is tropolone.
12. The method of claim 9 , wherein the topoisomerase inhibitor is topotecan.