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Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells
Date Issued
2012-11-29
Date Available
2013-11-29T10:10:30Z
Abstract
Introduction: In recent years much progress has been made in the development of tools for systems biology to study the levels of mRNA and protein, and their interactions within cells. However, few multiplexed methodologies are available to study cell signalling directly at the transcription factor level. 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.
Other Sponsorship
Engineering and Physical Sciences Research Council (EPSRC) Basic Technology Research Grant [grant number EP/E032745/1]. WK was also supported by Science Foundation Ireland [grant number 06/CE/B1129].
Type of Material
Journal Article
Publisher
Public Library of Science
Journal
PLoS ONE
Volume
7
Issue
11
Start Page
e50521
Copyright (Published Version)
2012 Public Library of Science
Language
English
Status of Item
Peer reviewed
This item is made available under a Creative Commons License
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