[Cellular test systems for the search for transcription factor activity modulators]

Area of Science:

• Molecular Biology
• Biochemistry
• Pharmacology

Context:

• Transcription factors (TFs) regulate gene expression and are key biotargets.
• Developing robust assay systems is crucial for identifying TF modulators.
• Existing methods may not fully capture the complex regulatory roles of TFs.

Purpose:

• To design and validate novel reporter assay systems for monitoring the activity of multiple transcription factors.
• To assess the functional activity of these reporter systems using transient transfection and specific inducers.
• To evaluate the efficacy of the developed systems by testing the known effects of aspirin and discovering new ones.

Summary:

• Luciferase reporter constructs were engineered with binding sites for CREB, NFAT, NF-kB, p53, STAT1, GAS, VDR, HSF1, and HIF1alpha.
• HEK293 cells were transiently transfected and treated with inducers to confirm functional activity via luciferase expression.
• Aspirin treatment revealed suppression of NF-kB, HIF1alpha, GAS, VDR, and HSF binding activity, with novel effects on HIF1alpha, GAS, VDR, and CREB.

Impact:

• Provides validated test systems for high-throughput screening of compounds targeting various transcription factors.
• Identifies novel inhibitory effects of aspirin on HIF1alpha, GAS, VDR, and CREB, expanding our understanding of its mechanisms.
• Facilitates drug discovery and mechanistic studies by offering a versatile platform for TF activity assessment.