Claims
- 1. A method for identifying a compound affecting the MAPK pathway comprising the steps of:
a) providing a cell stably transfected with a recombinant construct, said recombinant construct comprising a polynucleotide encoding a reporter gene, wherein said reporter gene operatively linked to the c-fos promoter; b) contacting said cell with a compound; and c) detecting a change in expression of said reporter gene; whereby a compound affecting the MAPK pathway is identified by detecting the change in expression of said reporter gene under control of the c-fos promoter.
- 2. A method of claim 1, wherein said reporter gene is the luciferase gene.
- 3. A method of claim 1 or 2, wherein said compound inhibits the MAPK pathway.
- 4. A method of claim 1 or 2, wherein said compound activates the MAPK pathway.
- 5. A method of claim 3, wherein said cell constitutively expresses low levels of invasion-associated genes, whereby stimulation of said invasion-associated genes occurs via activation of the MAPK pathway.
- 6. A method of claim 4, wherein said cell constitutively expresses low levels of invasion-associated genes, whereby stimulation of said invasion-associated genes occurs via activation of the MAPK pathway.
- 7. A method of claim 3, wherein said cell is weakly tumorigenic, whereby c-ets-1 mRNA expression is activated exclusively via the MAPK pathway in said cell.
- 8. A method of claim 4, wherein said cell is weakly tumorigenic, whereby c-ets-1 mRNA expression is activated exclusively via the MAPK pathway in said cell.
- 9. A method of claim 3, wherein said cell is a SNB-19 glioma cell.
- 10. A method of claim 4, wherein said cell is a SNB-19 glioma cell.
- 11. A method of claim 1, wherein the change in expression of said reporter gene is detected as a change in said reporter gene mRNA expression.
- 12. A method of claim 1, wherein the change in expression of said reporter gene is detected as a change in said reporter gene protein expression.
- 13. A method of claim 1, wherein the change in expression of said reporter gene is detected as a change in said reporter gene enzyme activity.
- 14. The method of claim 1, wherein the time course of the change in expression of said reporter gene corresponds to the time course of a change in c-fos gene expression.
- 15. The method of claim 1, wherein said recombinant construct further comprises at least a portion of the c-fos 3′-untranslated region sequence downstream of said reporter gene sufficient to cause the change in reporter gene expression to correspond to the change in the expression of c-fos.
- 16. A cell comprising a recombinant construct responsive to modulators of the MAPK pathway, wherein said recombinant construct comprises a polynucleotide encoding a reporter gene operatively linked to the c-fos promoter.
- 17. A cell of claim 16, wherein said reporter gene is the luciferase gene.
- 18. A cell of claim 16 or 17, wherein said recombinant construct is responsive to inhibitors of the MAPK pathway.
- 19. A cell of claim 16 or 17, wherein said recombinant construct is responsive to activators of the MAPK pathway.
- 20. A cell of claim 16 or 17, wherein said cell constitutively expresses low levels of invasion-associated genes, whereby stimulation of said invasion-associated genes occurs via activation of the MAPK pathway.
- 21. A cell of claim 16 or 17, wherein said cell is weakly tumorigenic, whereby c-ets-1 mRNA expression is activated exclusively via the MAPK pathway in said cell.
- 22. A cell of claim 16 or 17, wherein said cell is a SNB-19 glioma cell.
- 23. A cell of claim 16 or 17, wherein the time course of a change of expression of said reporter gene corresponds to the time course of a change in c-fos expression.
- 24. A cell of claim 16 or 17, wherein said recombinant construct further comprises at least a portion of the c-fos 3′-untranslated region sequence downstream of said reporter gene sufficient to cause reporter gene expression to correspond to the expression of c-fos.
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional App. No. 60/255,548 filed Dec. 14, 2000, which is incorporated by reference herein in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60255548 |
Dec 2000 |
US |