Parallel tracking of cAMP and PKA signaling dynamics in living cells with FRET-based fluorescent biosensors†
Abstract
Proper regulation of cellular functions relies upon a network of intricately interwoven signaling cascades in which multiple components must be tightly coordinated both spatially and temporally. To better understand how this network operates within the cellular environment, it is important to define the parameters of various signaling activities and to reveal the characteristic activity structure of the signaling cascades. This task calls for molecular tools capable of parallelly tracking multiple activities in cellular time and space with high sensitivity and specificity. Here, we present new biosensors developed based on two conveniently co-imageable FRET pairs consisting of CFP–RFP and YFP–RFP, specifically Cerulean-mCherry and mVenus-mCherry, for parallel monitoring of PKA activity and