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首页> 外文期刊>Journal of Neuroscience Methods >Optimization of a cAMP response element signal pathway reporter system.
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Optimization of a cAMP response element signal pathway reporter system.

机译：cAMP反应元件信号通路报告系统的优化。

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A sensitive cAMP response element (CRE) reporter system is essential for studying the cAMP/protein kinase A/cAMP response element binding protein signal pathway. Here we have tested a few CRE promoters and found one with high sensitivity to external stimuli. Using this optimal CRE promoter and the enhanced green fluorescent protein as the reporter, we have established a CRE reporter cell line. This cell line can be used to study the signal pathway by fluorescent microscope, fluorescence-activated cell analysis and luciferase assay. This cell line's sensitivity to forskolin, using the technique of fluorescence-activated cell sorting, was increased to approximately seven times that of its parental HEK 293 cell line, which is currently the most commonly used cell line in the field for the signal pathway study. Therefore, this newly created cell line is potentially useful for studying the signal pathway's modulators, which generally have weaker effect than its mediators. Our research has also established a general procedure for optimizing transcription-based reporter cell lines, which might be useful in performing the same task when studying many other transcription-based signal pathways.
机译：敏感的cAMP反应元件（CRE）报告系统对于研究cAMP /蛋白激酶A / cAMP反应元件结合蛋白信号通路至关重要。在这里，我们测试了一些CRE启动子，并发现了一个对外部刺激高度敏感的启动子。使用这种最佳的CRE启动子和增强的绿色荧光蛋白作为报告基因，我们建立了CRE报告基因细胞系。该细胞系可用于通过荧光显微镜，荧光激活的细胞分析和荧光素酶测定法研究信号途径。使用荧光激活细胞分选技术，该细胞系对毛喉素的敏感性提高到其亲代HEK 293细胞系的约七倍，后者是目前信号通路研究领域中最常用的细胞系。因此，这种新创建的细胞系潜在地可用于研究信号通路的调节剂，其作用通常比其介体弱。我们的研究还建立了优化基于转录的报告基因细胞系的通用程序，这在研究许多其他基于转录的信号途径时可能对执行相同的任务很有用。

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