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US 7,407,745 B1Granted
Method for screening anticancer agent
AssigneeChugai Seiyaku Kabushiki Kaisha
InventorsYoshihiro Sowa, Tetsuro Orita
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Abstract
By analyzing the mechanism for inducing a histone deacetylase inhibitor (Trichostatin A)-mediated expression of cyclin-CDK inhibitory factor having a proliferation suppressing effect (tumor-suppressing effect), it was revealed that the binding of Sp3 to the Sp1 binding sequence within a promoter is important in the expression of the above factor. Thus, a novel antitumor agent can be developed by screening a pharmaceutical agent capable of elevating Sp3 activity.
Claims10
A method of identifying an agent that activates TSA-responsive Sp3-mediated transcription, the method comprising:
- providing a mammalian cell in vitro having
- (a) a first vector comprising a first regulatory sequence operably linked to a nucleic acid sequence encoding a fusion protein, wherein the fusion protein comprises:
- (i) a fragment of human Sp3 (1) having transcriptional activation activity, and (2) comprising at least one glutamine rich region of a TSA responsive domain of human Sp3, wherein at least amino acids 495-517, 525-547, and 555-575 of the Sp3 Zinc finger region are lacking from said fragment of human Sp3, and
- (ii) a DNA binding domain of a heterologous protein;
- and
- (b) a second vector comprising a target binding sequence for the DNA binding domain of the fusion protein operably linked to a reporter gene;
- contacting the cell with a test agent;
- and
- selecting a test agent that increases the expression of the reporter gene compared to a control.
6 more elements
Dependencies are inferred from the claim text.