Methods - Here we describe a sensitive, plasmid-based RNA reporter methodology to study transcription factor activation in mammalian cells, and apply this technology to profiling 60 transcription factors in parallel. The methodology uses two robust and easily accessible detection platforms; quantitative real-time PCR for quantitative analysis and DNA microarrays for parallel, higher throughput analysis.
Findings - We test the specificity of the detection platforms with ten inducers and independently validate the transcription factor activation.
Conclusions - We report a methodology for the multiplexed study of transcription factor activation in mammalian cells that is direct and not theoretically limited by the number of available reporters.
Bibliographical note© 2012 Jiwaji et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- real-time polymerase chain reaction
- Western blotting
- transcription factors
- oligonucleotide array sequence analysis
- systems biology
- cell line
- cadmium chloride