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- W4306756089 abstract "ABSTRACT Cyclic diguanylate (c-di-GMP) is a ubiquitous second messenger that regulates a wide range of biological processes in bacteria, including motility, surface attachment, virulence and persistence. The regulatory networks controlling c-di-GMP are generally complex and understudied. This is largely due to a lack of appropriate tools to monitor dynamic changes of c-di-GMP concentrations in vivo in a non-invasive manner. Here, we develop a genetically-encoded ratiometric c-di-GMP biosensor, called cdGreen2, by applying a powerful directed evolution approach based on iterative fluorescence-activated cell sorting (FACS) under alternating c-di-GMP regimes. We demonstrate that cdGreen2 can robustly track c-di-GMP dynamics in live cells with ultra-high temporal resolution over multiple generations. To validate its exquisite diagnostic power, we utilize cdGreen2 to dissect the regulatory networks driving bimodal developmental programs in the environmental model organism Caulobacter crescentus and the human pathogen Pseudomonas aeruginosa . These studies disclose the molecular determinants governing cell cycle-dependent c-di-GMP oscillations in C. crescentus and surface-induced c-di-GMP asymmetry in P. aeruginosa . The sensitivity and versatility of cdGreen2 will help unveil c-di-GMP dynamics in a wide range of organisms with unprecedented temporal resolution. The simple, yet powerful design principles underlying cdGreen2 will serve as a blueprint for the development of similar, orthogonal biosensors for other signaling molecules, metabolites or antibiotics, paving the way to uncover the complex interplay of small molecule-based networks with unprecedented spatiotemporal resolution." @default.
- W4306756089 created "2022-10-19" @default.
- W4306756089 creator A5010461994 @default.
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- W4306756089 date "2022-10-18" @default.
- W4306756089 modified "2023-10-16" @default.
- W4306756089 title "A Novel Biosensor Reveals Dynamic Changes of C-di-GMP in Differentiating Cells with Ultra-High Temporal Resolution" @default.
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- W4306756089 doi "https://doi.org/10.1101/2022.10.18.512705" @default.
- W4306756089 hasPublicationYear "2022" @default.
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