Claims
- 1. A method for identifying a compound that modulates the hypocretin system comprising contacting a test compound to cells equipped with the 5′ flanking promoter of the preprohypocretin gene operably linked to a nucleic acid sequence and determining whether the test compound alters transcription of the nucleic acid sequence in the cells, wherein the test compound's ability to alter transcription is indicative of a compound that modulates the hypocretin system.
- 2. The method of claim 1, wherein the cells are naturally occurring preprohypocretin-expressing cells.
- 3. The method of claim 2, wherein the naturally occurring preprohypocretin cells are isolated from the posterior hypothalamus, bladder tissue, or tissue of the gastrointestinal tract.
- 4. The method of claim 1, wherein the cells are genetically manipulated to be equipped with the 5′ flanking promoter of the preprohypocretin gene.
- 5. The method of claim 4, wherein the genetically manipulated cells are selected from the group consisting of neuroblastoma cells, kidney cells, and ovary cells.
- 6. The method of claim 5, wherein the nucleic acid sequence codes for a known gene.
- 7. The method of claim 6, wherein alteration of transcription is evidenced by a change in expression of the gene when compared to expression of the gene without the compound.
- 8. The method of claim 7, wherein the compound enhances expression of the gene.
- 9. The method of claim 8, wherein the compound binds to the 5′-flanking promoter of the preprohypocretin gene.
- 10. The method of claim 7, wherein the compound decreases expression of the gene.
- 11. The method of claim 10, wherein the compound binds to the 5′-flanking promoter of the preprohypocretin gene.
- 12. The method of claim 1, which is carried out in vitro.
- 13. An isolated DNA fragment coding for the 5′ flanking promoter of the preprohypocretin gene.
- 14. The isolated DNA fragment of claim 13, comprising the nucleotide sequence of SEQ ID NO: 1.
- 15. An expression vector comprising the DNA fragment of claim 13.
- 16. The expression vector of claim 15, wherein the DNA fragment comprises the nucleotide sequence of SEQ ID NO: 1.
- 17. A host cell transformed with the expression vector of claim 15.
- 18. A compound that modulates the hypocretin system, wherein the compound is identified by the steps comprising contacting a preprohypocretin-expressing cell with the compound and determining whether the compound alters preprohypocretin expression in the preprohypocretin-expressing cell, wherein the compound's ability to alter preprohypocretin expression is indicative of a compound that modulates the hypocretin system.
- 19. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 18.
- 20. The composition of claim 19, further comprising a pharmaceutically acceptable carrier.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation application of U.S. patent application Ser. No. 10/029,427, filed on Dec. 19, 2001, which claims priority under 35 U.S.C. §119(e)(1) to U.S. Provisional Application Serial No. 60/258,069, filed Dec. 20, 2000, all of which are incorporated by reference in their entireties herein.
REFERENCE TO GOVERNMENT SUPPORT
[0002] This invention was funded in part by the National Institute of Mental Health under NIH Grant Nos. 1 R01MH61755 and 1 R01HL/MH59658. The United States Government has certain rights in this invention.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60258069 |
Dec 2000 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
10029427 |
Dec 2001 |
US |
Child |
10830638 |
Apr 2004 |
US |