Unique reporter-based sensor platforms to monitor signalling in cells

Jiwaji, M. et al. (2012) Unique reporter-based sensor platforms to monitor signalling in cells. PLoS ONE, 7(11), e50521. (doi:10.1371/journal.pone.0050521)

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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. <p/>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. <p/>Findings: We test the specificity of the detection platforms with ten inducers and independently validate the transcription factor activation. <p/>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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:West, Dr Adam and Cumming, Dr Sarah and Pitt, Dr Andrew and West, Dr Katherine and Barkess, Dr Grainne and Gibriel, Dr Abdullah and Girolami, Prof Mark and Daly, Dr Ronan
Authors: Jiwaji, M., Daly, R., Gibriel, A., Barkess, G., McLean, P., Yang, J., Pansare, K./, Cumming, S., McLauchlan, A., Kamola, P., Bhutta, M., West, A., West, K., Kolch, W., Girolami, M., and Pitt, A.
Subjects:Q Science > Q Science (General)
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Science and Engineering > School of Computing Science
Journal Name:PLoS ONE
Publisher:Public Library of Science
ISSN:1932-6203
ISSN (Online):1932-6203
Published Online:29 November 2012
Copyright Holders:Copyright © 2012 The Authors
First Published:First published in PLoS ONE 7(11):e50521
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
436881The molecular noseAndrew PittEngineering & Physical Sciences Research Council (EPSRC)EP/E032745/1RI MOLECULAR CELL & SYSTEMS BIOLOGY