Patent ID: 8592147
Filing Date: 2013-11-26
Classification: C12Q,G01N

Abstract:
1. A method for identifying a small molecule test compound that inhibits a promoter activated by cell cycle inhibition comprising: (a) providing cells comprising a first inducible promoter linked to a gene that encodes a protein that directly or indirectly inhibits cell cycle progression and a recombinant expression vector comprising a reporter gene encoding a detectable protein under transcriptional control of a second promoter that is activated by cell cycle inhibition; (b) dividing the cells into four aliquots; (c) treating the first aliquot with a small molecule test compound, wherein said first promoter is activated; (d) treating the second aliquot with the small molecule test compound, wherein said first promoter is not activated; (e) providing the third aliquot, wherein the third aliquot is not treated with the small molecule test compound and wherein said first promoter is activated; (f) providing the fourth aliquot, wherein the fourth aliquot is not treated with the small molecule test compound and wherein said first promoter is not activated; (g) determining total cell number of each aliquot; (h) determining the amount of the detectable protein in each aliquot; (i) calculating the ratio of detectable protein to the total cell number for each aliquot; and (j) comparing the ratio of each aliquot, wherein the small molecule test compound is identified as a small molecule compound that inhibits a promoter activated by cell cycle inhibition if the ratio from the third aliquot is higher than the ratio of the first aliquot, which is higher than the ratio from the fourth aliquot, which is equal to or higher than the ratio from the second aliquot.