Unique reporter-based sensor platforms to monitor signalling in cells

Meesbah Jiwaji; Rónán Daly; Abdullah Gibriel; Gráinne Barkess; Pauline McLean; Jingli Yang; Kshama Pansare; Sarah A. Cumming; Alisha McLauchlan; Piotr J. Kamola; Musab S. Bhutta; Adam G. West; Katherine L. West; Walter Kolch; Mark A. Girolami; Andrew R. Pitt

doi:10.1371/journal.pone.0050521

Unique reporter-based sensor platforms to monitor signalling in cells

Meesbah Jiwaji, Rónán Daly, Abdullah Gibriel, Gráinne Barkess, Pauline McLean, Jingli Yang, Kshama Pansare, Sarah A. Cumming, Alisha McLauchlan, Piotr J. Kamola, Musab S. Bhutta, Adam G. West, Katherine L. West, Walter Kolch, Mark A. Girolami, Andrew R. Pitt

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	e50521
Number of pages	10
Journal	PLoS ONE
Volume	7
Issue number	11
DOIs	https://doi.org/10.1371/journal.pone.0050521
Publication status	Published - 29 Nov 2012

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.

Keywords

real-time polymerase chain reaction
dexamethasone
Western blotting
transcription factors
oligonucleotide array sequence analysis
humans
systems biology
plasmids
cell line
cadmium chloride
forskolin

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10.1371/journal.pone.0050521

Unique reporter-based sensor platforms to monitor signalling in cells
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.
Submitted manuscript, 1.2 MBLicence: CC BY 3.0

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0050521

Cite this

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title = "Unique reporter-based sensor platforms to monitor signalling in cells",

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.",

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AU - McLean, Pauline

AU - Yang, Jingli

AU - Pansare, Kshama

AU - Cumming, Sarah A.

AU - McLauchlan, Alisha

AU - Kamola, Piotr J.

AU - Bhutta, Musab S.

AU - West, Adam G.

AU - West, Katherine L.

AU - Kolch, Walter

AU - Girolami, Mark A.

AU - Pitt, Andrew R.

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N2 - 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.

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Unique reporter-based sensor platforms to monitor signalling in cells

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Keywords

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