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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 l...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3510088/ https://www.ncbi.nlm.nih.gov/pubmed/23209767 http://dx.doi.org/10.1371/journal.pone.0050521 |
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author | 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 A. Pitt, Andrew R. |
author_facet | 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 A. Pitt, Andrew R. |
author_sort | Jiwaji, Meesbah |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3510088 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35100882012-12-03 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 McLauchlan, Alisha Kamola, Piotr J. Bhutta, Musab S. West, Adam G. West, Katherine L. Kolch, Walter Girolami, Mark A. Pitt, Andrew R. PLoS One Research Article 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. Public Library of Science 2012-11-29 /pmc/articles/PMC3510088/ /pubmed/23209767 http://dx.doi.org/10.1371/journal.pone.0050521 Text en © 2012 Jiwaji et al http://creativecommons.org/licenses/by/4.0/ 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 properly credited. |
spellingShingle | Research Article 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 A. Pitt, Andrew R. Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title | Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title_full | Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title_fullStr | Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title_full_unstemmed | Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title_short | Unique Reporter-Based Sensor Platforms to Monitor Signalling in Cells |
title_sort | unique reporter-based sensor platforms to monitor signalling in cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3510088/ https://www.ncbi.nlm.nih.gov/pubmed/23209767 http://dx.doi.org/10.1371/journal.pone.0050521 |
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