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. and Pitt, Andrew R. (2012). Unique reporter-based sensor platforms to monitor signalling in cells. PLoS ONE, 7 (11),
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.
Publication DOI: | https://doi.org/10.1371/journal.pone.0050521 |
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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,General Agricultural and Biological Sciences,General Biochemistry,Genetics and Molecular Biology,General Medicine |
Publication ISSN: | 1932-6203 |
Last Modified: | 30 Sep 2024 10:14 |
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. ( 0000-0003-3619-6503) |