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A BW Reporter System for Studying Receptor-Ligand Interactions
Published on: January 7, 2019
Reporter Gene Assay for Monitoring BCL6 Reactivation
Catherine L Ingram1, Radosław P Nowak2, Thomas M Geiger3
1Institute of Structural Biology, University of Bonn, Bonn, Germany.
Abstract:
Transcription factors are an exciting class of drug targets. Recent developments in small molecule-induced proximity allow for small molecule control of their activity by recruitment of epigenetic proteins. In this chapter, we describe the development of a transcriptional activity assay for the transcription factor BCL6 with a small molecule activator, TCIP1, in HEK293T cells [1]. The assay is based on the Bio-Glo Luciferase assay system and uses transient transfection of a BCL6-expressing plasmid, as well as a reporter plasmid where a BCL6 DNA-binding sequence controls expression of a luciferase protein. The method described here is specific to BCL6 but can easily be optimized to monitor the activity of other transcription factors by altering the DNA-binding sequence of the reporter.
Insights
Researchers developed a new assay to measure the activity of the BCL6 transcription factor using a small molecule activator. This method, utilizing luciferase reporter gene technology, offers a versatile platform for studying transcription factor drug targets.
Area of Science:
- Molecular Biology
- Drug Discovery
- Epigenetics
Background:
- Transcription factors are key regulators of gene expression and emerging drug targets.
- Small molecule-induced proximity is a novel strategy to control transcription factor activity.
- BCL6 is a significant transcription factor implicated in various cellular processes.
Purpose of the Study:
- To develop and validate a transcriptional activity assay for the BCL6 transcription factor.
- To demonstrate the utility of small molecule activators in modulating transcription factor function.
- To establish a flexible assay adaptable for other transcription factors.
Main Methods:
- Development of a luciferase reporter assay system in HEK293T cells.
- Utilized transient transfection with BCL6-expressing and reporter plasmids.
- Employed a specific BCL6 DNA-binding sequence to control luciferase expression.
Main Results:
- Successfully established a functional assay to measure BCL6 transcriptional activity.
- Demonstrated the ability of the small molecule activator TCIP1 to modulate BCL6 activity.
- Validated the specificity of the assay for BCL6.
Conclusions:
- The developed assay provides a robust method for studying BCL6 activity.
- This platform enables the screening and characterization of small molecules targeting transcription factors.
- The assay's adaptability facilitates research into diverse transcription factor drug targets.
