Related Experiment Videos
A high-throughput mammalian cell-based transient transfection assay
Daniel J Noonan1, Kenneth Henry, Michelle L Twaroski
1Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 3, 2004
Summary
This study presents a high-throughput method for assaying transcription factor activity in mammalian cells. The cost-effective, 96-well plate protocol enables efficient screening of gene regulatory factors.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Gene expression in eukaryotes is regulated by transcription factors that target coregulatory complexes.
- Understanding transcription factor activity is crucial for both academic and industrial research.
- Various methods exist for assaying transcription factor activity in mammalian cells.
Purpose of the Study:
- To develop a high-throughput methodology for screening transcription factor activities.
- To adapt existing methods for broader applicability in academic laboratories.
- To provide a cost-effective solution for studying gene regulation.
Main Methods:
- Utilizes co-transfection of mammalian expression and luciferase reporter plasmids.
- Employs a 96-well plate format for high-throughput screening.
- Focuses on reconstituting transcription events in mammalian host cells.
Main Results:
- A high-throughput version of a transcription factor assay methodology is presented.
- The method is adaptable for screening a variety of transcription factors.
- Detailed protocols and cost-effective measures are provided for laboratory implementation.
Conclusions:
- The developed high-throughput assay facilitates efficient screening of transcription factors.
- This cost-effective technology is suitable for low-budget academic laboratories.
- The methodology aids in understanding the regulatory forces of gene expression.