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Droplet Digital TRAP ddTRAP: Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction
Published on: May 3, 2019
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Promoter trapping method: transcription factor purification using human telomerase reverse transcriptase promoter.
Linda I Nagore1, YanWen Zhou1, Robert J Nadeau1
1Department of Chemistry, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249 USA.
Proteome Science
|November 27, 2014
Summary
This study uses promoter trapping to identify transcription factors regulating the telomerase gene, crucial for cancer. The method successfully isolated known factors like AP-2 and SP1, offering potential new cancer treatment strategies.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Background:
- Transcription factors regulate gene activity by binding to promoter regions.
- Identifying low-abundance transcription factors is challenging.
- Understanding gene regulation is key to developing new cancer treatments.
Purpose of the Study:
- To develop and validate a purification technique for transcription factors.
- To study the telomerase promoter, which is highly active in cancer cells.
- To explore potential new cancer treatment strategies by controlling telomerase transcription.
Main Methods:
- Utilized promoter trapping with column chromatography to isolate proteins.
- Analyzed trapped complexes using liquid chromatography tandem mass spectrometry.
- Focused on the telomerase promoter region (-170 - +91).
Main Results:
- Successfully isolated a transcriptionally active and reproducible complex from the telomerase promoter.
- Identified known transcription factors AP-2 and SP1.
- Demonstrated that AP-2 and SP1 can bind to each other's response elements.
Conclusions:
- Verified the efficacy and versatility of promoter trapping for identifying transcription factors.
- Confirmed the ability of promoter trapping to identify known regulators of telomerase transcription.
- Validated promoter trapping as a reliable method for studying gene regulation.
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