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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Abstract	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.
AbstractList	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. 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. [PUBLICATION ABSTRACT] 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.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.
Audience	Academic
Author	Gambarian, Maria Diatchenko, Luda Medvedev, Alexander Romanov, Sergei Makarov, Sergei Poltoratskaya, Natalia Medvedeva, Liubov Moeser, Matt Gambarian, Mikhail
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BackLink	https://www.ncbi.nlm.nih.gov/pubmed/18297081$$D View this record in MEDLINE/PubMed
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Snippet	We developed a high-content reporter system that allows quantitative assessment of activities of multiple transcription factors (TFs) in a eukaryotic cell. The...
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SubjectTerms	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
Title	Homogeneous reporter system enables quantitative functional assessment of multiple transcription factors
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