Homogeneous reporter system enables quantitative functional assessment of multiple transcription factors
Homogeneous reporter system enables quantitative functional assessment of multiple transcription factors
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DOI：
10.1038/nmeth.1186
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发表时间：
2008-03-01
期刊：
影响因子：
48
通讯作者：
Makarov, Sergei
中科院分区：
文献类型：
--
作者：
Romanov, Sergei;Medvedev, Alexander;Makarov, Sergei
我们开发了一个高含量的报告系统，可以定量评估真核细胞中多种转录因子（tf）的活性。该系统包括一个报告结构库，根据其转录率进行评估。所有的报告者都会产生本质上相同的信息，这些信息都要经过“处理”，从而产生一系列可区分的片段，并进行定量分析。报告者库的同质性为所有报告者提供了本质上统一的检测条件，并提供了评估的可重复性、准确性和稳健性。我们证明，该技术可用于识别传递细胞对诱导剂反应的途径，并且使用该系统产生的TF活性谱代表了一个稳定和持续的细胞特征，可以清楚地区分不同的细胞类型和病理状况。该技术通过分析多个tf的活动，为信号转导网络的功能表征提供了一个框架。
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.