Unique reporter-based sensor platforms to monitor signalling in cells


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.

Publication DOI: https://doi.org/10.1371/journal.pone.0050521
Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > Aston Pharmacy School
College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
Additional Information: © 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.
Uncontrolled Keywords: real-time polymerase chain reaction,dexamethasone,Western blotting,transcription factors,oligonucleotide array sequence analysis,humans,systems biology,plasmids,cell line,cadmium chloride,forskolin,Agricultural and Biological Sciences(all),Biochemistry, Genetics and Molecular Biology(all),Medicine(all)
Publication ISSN: 1932-6203
Last Modified: 04 Jan 2024 08:14
Date Deposited: 12 Jun 2013 08:36
Full Text Link: http://www.plos ... al.pone.0050521
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2012-11-29
Authors: Jiwaji, Meesbah
Daly, Rónán
Gibriel, Abdullah
Barkess, Gráinne
McLean, Pauline
Yang, Jingli
Pansare, Kshama
Cumming, Sarah A.
McLauchlan, Alisha
Kamola, Piotr J.
Bhutta, Musab S.
West, Adam G.
West, Katherine L.
Kolch, Walter
Girolami, Mark A.
Pitt, Andrew R. (ORCID Profile 0000-0003-3619-6503)



Version: Draft Version

License: Creative Commons Attribution

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