Protein kinases play pivotal roles in intracellular signal transduction, and dysregulation of kinases leads to pathological results such as malignant tumors. Kinase activity has hitherto been measured by biochemical methods such as in vitro phosphorylation assay and western blotting. However, these methods are less useful to explore spatial and temporal changes in kinase activity and its cell-to-cell variation. Recent advances in fluorescent proteins and live-cell imaging techniques enable us to visualize kinase activity in living cells with high spatial and temporal resolutions. Several genetically encoded kinase activity reporters, which are based on the modes of action of kinase activation and phosphorylation, are currently available. These reporters are classified into single-fluorophore kinase activity reporters and Förster (or fluorescence) resonance energy transfer (FRET)-based kinase activity reporters. Here, we introduce the principles of genetically encoded kinase activity reporters, and discuss the advantages and disadvantages of these reporters.Key words: kinase, FRET, phosphorylation, KTR.

中文翻译：

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遗传编码的蛋白激酶活性报告基因的活细胞成像。

蛋白激酶在细胞内信号转导中起关键作用，激酶的失调可导致病理结果，例如恶性肿瘤。迄今为止，激酶活性已经通过诸如体外磷酸化测定和蛋白质印迹的生化方法进行了测量。但是，这些方法对于探索激酶活性及其细胞间变化的时空变化不太有用。荧光蛋白和活细胞成像技术的最新进展使我们能够以高时空分辨率可视化活细胞中的激酶活性。目前有几种基于激酶激活和磷酸化作用方式的基因编码的激酶活性报告基因。这些报道分子分为单荧光团激酶活性报道分子和基于Förster（或荧光）共振能量转移（FRET）的激酶活性报道分子。在这里，我们介绍了遗传编码激酶活性报告基因的原理，并讨论了这些报告基因的优缺点。关键词：激酶，FRET，磷酸化，KTR