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Unique reporter-based sensor platforms to monitor signalling in cells
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Jiwaji, Meesbah, Daly, Rónán, Gibriel, Abdullah, Barkess, Gráinne, McLean, Pauline, Yang, Jingli, Pansare, Kshama, Cumming, Sarah, McLauchlan, Alisha, Kamola, Piotr J., Bhutta, Musab S., West, Adam G., West, Katherine L., Kolch, Walter, Girolami, Mark and Pitt, A. R. (2012) Unique reporter-based sensor platforms to monitor signalling in cells. PLoS One, Volume 7 (Number 11). Article number e50521. doi:10.1371/journal.pone.0050521 ISSN 1932-6203.
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Official URL: http://dx.doi.org/10.1371/journal.pone.0050521
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.
| Item Type: | Journal Article | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| Subjects: | Q Science > QP Physiology | ||||||||
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Statistics | ||||||||
| Library of Congress Subject Headings (LCSH): | Messenger RNA, RNA-protein interactions | ||||||||
| Journal or Publication Title: | PLoS One | ||||||||
| Publisher: | Public Library of Science | ||||||||
| ISSN: | 1932-6203 | ||||||||
| Official Date: | 29 November 2012 | ||||||||
| Dates: |
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| Volume: | Volume 7 | ||||||||
| Number: | Number 11 | ||||||||
| Article Number: | Article number e50521 | ||||||||
| DOI: | 10.1371/journal.pone.0050521 | ||||||||
| Status: | Peer Reviewed | ||||||||
| Publication Status: | Published | ||||||||
| Access rights to Published version: | Open Access (Creative Commons) | ||||||||
| Date of first compliant deposit: | 28 December 2015 | ||||||||
| Date of first compliant Open Access: | 28 December 2015 | ||||||||
| Funder: | Engineering and Physical Sciences Research Council (EPSRC), Science Foundation Ireland (SFI) | ||||||||
| Grant number: | EP/ E032745/1 (EPSRC), 06/CE/B1129 (SFI) |
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