Unique reporter-based sensor platforms to monitor signalling in cells

独特的基于报告器的传感器平台可监测细胞内的信号传导

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作者: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

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.

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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