Methods for identifying compounds which modulate circadian rhythm
AssigneeThe General Hospital Corporation
InventorsSteven M. Reppert, David R. Weaver, Mark Zylka, Xiaowei Jin, Kazuhiko Kume, Sriram Sathyanarayanan, Lauren Shearman
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Abstract
The invention is based, in part, on the discovery that the CRY proteins and the PER2 protein function as important modulators of mammalian circadian rhythm. The invention includes methods of modulating the circadian rhythm and identifying compounds that modulate the circadian rhythm.
Claims33
A method for identifying a compound which binds to a mammalian CRY protein, the method comprising:
- contacting the CRY protein with a test compound;
- and
- determining whether the CRY protein binds to the test compound, wherein binding by the test compound to the CRY protein indicates that the test compound is a CRY protein binding compound.
A method for identifying a compound which disrupts the association of a CRY protein and a PER protein, the method comprising:
- contacting a test compound with the CRY protein in the presence of the PER protein;
- and
- determining whether the test compound disrupts the association of the CRY protein and the PER protein, wherein a decrease in the association in the presence of the test compound compared to the association in the absence of the test compound indicates that the test compound disrupts the association of the CRY protein and the PER protein.
A method for identifying a compound which disrupts the association of a CRY protein and a TIM protein, the method comprising:
- contacting a test compound with the CRY protein in the presence of the TIM protein;
- and
- determining whether the test compound disrupts the association of the CRY protein and the TIM protein, wherein a decrease in the association in the presence of the test compound compared to the association in the absence of the test compound indicates that the test compound disrupts the association of the CRY protein and the TIM protein.
The method of identifying a compound that disrupts the association between a CRY protein and a CLOCK:BMAL-1 complex, the method comprising:
- contacting a test compound with the CRY protein in the presence of a CLOCK protein amd a BMAL-1 protein;
- and
- determining whether the test compound disrupts the association of the CRY protein with a complex of the CLOCK protein and the BMAL-1 protein, wherein a decrease in the association in the presence of the test compound compared to the association in the absence of the test compound indicates that the test compound disrupts the association of the CRY protein and the CLOCK:BMAL-1 complex.
A method for identifying a compound which disrupts the association of a CRY protein and a BMAL-1 protein, the method comprising:
- contacting a test compound with the CRY protein in the presence of the BMAL-1 protein;
- and
- determining whether the test compound disrupts the association of the CRY protein and the BMAL-1 protein, wherein a decrease in the association in the presence of the test compound compared to the association in the absence of the test compound indicates that the test compound disrupts the association of the CRY protein and the BMAL-1 protein.
A method for identifying a compound which disrupts the association of a first CRY protein and a second CRY protein, the method comprising:
- contacting a test compound with the first and second CRY proteins;
- and
- determining whether the test compound disrupts the association of the first CRY protein with the second CRY protein, wherein the second CRY protein has an amino acid sequence the same as or different than the first CRY protein, and wherein a decrease in the association in the presence of the test compound compared to the association in the absence of the test compound indicates that the test compound disrupts the association of the first protein with the second CRY protein.
A method for identifying a compound that blocks CRY induced-inhibition of CLOCK:BMAL-1 transcription in a cell, the method comprising:
- providing a cell comprising a CRY protein, a CLOCK:BMAL-1 complex, and a DNA comprising an E-box operatively linked to a reporter gene;
- introducing the compound into the cell;
- and
- assaying for transcription of the reporter gene in the cell, wherein an increase in transcription in the presence of the compound compared to transcription in the absence of the compound indicates that the compound blocks CRY-induced inhibition of CLOCK:BMAL-1-mediated transcription in a cell.
Dependencies are inferred from the claim text.