Homogeneous reporter system enables quantitative functional assessment of multiple transcription factors

We developed a high-content reporter system that allows quantitative assessment of activities of multiple transcription factors (TFs) in a eukaryotic cell. The system comprises a library of reporter constructs that are evaluated according to their transcription rates. All reporters produce essential...

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Vydáno v:Nature methods Ročník 5; číslo 3; s. 253 - 260
Hlavní autoři: Romanov, Sergei, Medvedev, Alexander, Gambarian, Maria, Poltoratskaya, Natalia, Moeser, Matt, Medvedeva, Liubov, Gambarian, Mikhail, Diatchenko, Luda, Makarov, Sergei
Médium: Journal Article
Jazyk:angličtina
Vydáno: New York Nature Publishing Group US 01.03.2008
Nature Publishing Group
Témata:
Animals
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedical research
Carbazoles - pharmacology
Cell Line, Tumor
Cellular biology
Cellular signal transduction
Colforsin - pharmacology
DNA binding proteins
Gene Expression Profiling - methods
Gene Library
Genes, Reporter - physiology
Genetic aspects
Genetic regulation
Homogeneity
Humans
Indoles - pharmacology
Life Sciences
Oligonucleotide Array Sequence Analysis - methods
Physiological aspects
Polymerase Chain Reaction
Proteomics
Rats
Receptors, Aryl Hydrocarbon - drug effects
Reproducibility of Results
Research methodology
Signal transduction
Transcription Factors - genetics
Transfection - methods
Transforming Growth Factor beta - pharmacology
United States
ISSN:1548-7091, 1548-7105, 1548-7105
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Shrnutí:We developed a high-content reporter system that allows quantitative assessment of activities of multiple transcription factors (TFs) in a eukaryotic cell. The system comprises a library of reporter constructs that are evaluated according to their transcription rates. All reporters produce essentially identical messages that are subjected to 'processing', which generates a spectrum of distinguishable fragments that are analyzed quantitatively. The homogeneity of the reporter library afforded inherently uniform detection conditions for all reporters and provided repeatability, accuracy and robustness of assessment. We showed that this technology can be used to identify pathways transmitting cell responses to inducers, and that the profile of TF activities generated using this system represents a stable and sustained cell signature that clearly distinguishes different cell types and pathological conditions. This technology provides a framework for functional characterization of signal transduction networks through profiling activities of multiple TFs.
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ISSN:1548-7091
1548-7105
1548-7105
DOI:10.1038/nmeth.1186