title: Unique reporter-based sensor platforms to monitor signalling in cells.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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.
authors.affiliation: Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Meesbah
authors.initials: M
authors.lastname: Jiwaji
authors.forename: Rónán
authors.initials: R
authors.lastname: Daly
authors.forename: Abdullah
authors.initials: A
authors.lastname: Gibriel
authors.forename: Gráinne
authors.initials: G
authors.lastname: Barkess
authors.forename: Pauline
authors.initials: P
authors.lastname: McLean
authors.forename: Jingli
authors.initials: J
authors.lastname: Yang
authors.forename: Kshama
authors.initials: K
authors.lastname: Pansare
authors.forename: Sarah
authors.initials: S
authors.lastname: Cumming
authors.forename: Alisha
authors.initials: A
authors.lastname: McLauchlan
authors.forename: Piotr J
authors.initials: PJ
authors.lastname: Kamola
authors.forename: Musab S
authors.initials: MS
authors.lastname: Bhutta
authors.forename: Adam G
authors.initials: AG
authors.lastname: West
authors.forename: Katherine L
authors.initials: KL
authors.lastname: West
authors.forename: Walter
authors.initials: W
authors.lastname: Kolch
authors.forename: Mark A
authors.initials: MA
authors.lastname: Girolami
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Pitt
mesh_terms: D015153:Blotting, Western; D019256:Cadmium Chloride / Q000494:pharmacology; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D049490:Systems Biology / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D014157:Transcription Factors; D005576:Colforsin; D003907:Dexamethasone; D019256:Cadmium Chloride