Unique reporter-based sensor platforms to monitor signalling in cells.
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.Here we d...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3510088?pdf=render |
| _version_ | 1811192985701842944 |
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| author | Meesbah Jiwaji Rónán Daly Abdullah Gibriel Gráinne Barkess Pauline McLean Jingli Yang Kshama Pansare Sarah Cumming Alisha McLauchlan Piotr J Kamola Musab S Bhutta Adam G West Katherine L West Walter Kolch Mark A Girolami Andrew R Pitt |
| author_facet | Meesbah Jiwaji Rónán Daly Abdullah Gibriel Gráinne Barkess Pauline McLean Jingli Yang Kshama Pansare Sarah Cumming Alisha McLauchlan Piotr J Kamola Musab S Bhutta Adam G West Katherine L West Walter Kolch Mark A Girolami Andrew R Pitt |
| author_sort | Meesbah Jiwaji |
| collection | DOAJ |
| description | 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.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.We test the specificity of the detection platforms with ten inducers and independently validate the transcription factor activation.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. |
| first_indexed | 2024-04-12T00:00:58Z |
| format | Article |
| id | doaj.art-b51b65aabb0b4effa3a0670b0f43cb99 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T00:00:58Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-b51b65aabb0b4effa3a0670b0f43cb992022-12-22T03:56:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01711e5052110.1371/journal.pone.0050521Unique reporter-based sensor platforms to monitor signalling in cells.Meesbah JiwajiRónán DalyAbdullah GibrielGráinne BarkessPauline McLeanJingli YangKshama PansareSarah CummingAlisha McLauchlanPiotr J KamolaMusab S BhuttaAdam G WestKatherine L WestWalter KolchMark A GirolamiAndrew R PittIn 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.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.We test the specificity of the detection platforms with ten inducers and independently validate the transcription factor activation.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.http://europepmc.org/articles/PMC3510088?pdf=render |
| spellingShingle | Meesbah Jiwaji Rónán Daly Abdullah Gibriel Gráinne Barkess Pauline McLean Jingli Yang Kshama Pansare Sarah Cumming Alisha McLauchlan Piotr J Kamola Musab S Bhutta Adam G West Katherine L West Walter Kolch Mark A Girolami Andrew R Pitt Unique reporter-based sensor platforms to monitor signalling in cells. PLoS ONE |
| 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 |
| url | http://europepmc.org/articles/PMC3510088?pdf=render |
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