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 journalArticle

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.
LanguageEnglish
Article numbere50521
Number of pages10
JournalPLoS ONE
Volume7
Issue number11
DOIs
Publication statusPublished - 29 Nov 2012

Fingerprint

Transcription Factors
transcription factors
monitoring
Transcriptional Activation
Sensors
Chemical activation
cells
Cells
Cell signaling
methodology
Systems Biology
Microarrays
Oligonucleotide Array Sequence Analysis
Cell Communication
quantitative analysis
Real-Time Polymerase Chain Reaction
plasmids
quantitative polymerase chain reaction
Plasmids
Throughput

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

Cite this

Jiwaji, M., Daly, R., Gibriel, A., Barkess, G., McLean, P., Yang, J., ... Pitt, A. R. (2012). Unique reporter-based sensor platforms to monitor signalling in cells. PLoS ONE, 7(11), [e50521]. https://doi.org/10.1371/journal.pone.0050521
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. / Unique reporter-based sensor platforms to monitor signalling in cells. In: PLoS ONE. 2012 ; Vol. 7, No. 11.
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Jiwaji, M, Daly, R, Gibriel, A, Barkess, G, McLean, P, Yang, J, Pansare, K, Cumming, SA, McLauchlan, A, Kamola, PJ, Bhutta, MS, West, AG, West, KL, Kolch, W, Girolami, MA & Pitt, AR 2012, 'Unique reporter-based sensor platforms to monitor signalling in cells' PLoS ONE, vol. 7, no. 11, e50521. https://doi.org/10.1371/journal.pone.0050521

Unique reporter-based sensor platforms to monitor signalling in cells. / 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.

In: PLoS ONE, Vol. 7, No. 11, e50521, 29.11.2012.

Research output: Contribution to journalArticle

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T1 - Unique reporter-based sensor platforms to monitor signalling in cells

AU - Jiwaji, Meesbah

AU - Daly, Rónán

AU - Gibriel, Abdullah

AU - Barkess, Gráinne

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.

N1 - © 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.

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Y1 - 2012/11/29

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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Jiwaji M, Daly R, Gibriel A, Barkess G, McLean P, Yang J et al. Unique reporter-based sensor platforms to monitor signalling in cells. PLoS ONE. 2012 Nov 29;7(11). e50521. https://doi.org/10.1371/journal.pone.0050521