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- W3089594507 abstract "Protein kinases control nearly every facet of cellular function. These key signaling nodes integrate diverse pathway inputs to regulate complex physiological processes, and aberrant kinase signaling is linked to numerous pathologies. While fluorescent protein-based biosensors have revolutionized the study of kinase signaling by allowing direct, spatiotemporally precise kinase activity measurements in living cells, powerful new molecular tools capable of robustly tracking kinase activity dynamics across diverse experimental contexts are needed to fully dissect the role of kinase signaling in physiology and disease. Here, we report the development of an ultrasensitive, second-generation excitation-ratiometric protein kinase A (PKA) activity reporter (ExRai-AKAR2), obtained via high-throughput linker library screening, that enables sensitive and rapid monitoring of live-cell PKA activity across multiple fluorescence detection modalities, including plate reading, cell sorting and one- or two-photon imaging. Notably, in vivo visual cortex imaging in awake mice reveals highly dynamic neuronal PKA activity rapidly recruited by forced locomotion. A fluorescence-based sensor of PKA activity has increased brightness, dynamic range and signal-to-noise ratio over related sensors and is useful for visualizing kinase activity in HeLa cells, primary neurons and the cortex of awake mice." @default.
- W3089594507 created "2020-10-08" @default.
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- W3089594507 date "2020-09-28" @default.
- W3089594507 modified "2023-10-15" @default.
- W3089594507 title "An ultrasensitive biosensor for high-resolution kinase activity imaging in awake mice" @default.
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- W3089594507 doi "https://doi.org/10.1038/s41589-020-00660-y" @default.
- W3089594507 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7773213" @default.
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