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J Cell Sci. 2014 Mar 15;127(Pt 6):1151-60. doi: 10.1242/jcs.099994.

Genetically encoded molecular probes to visualize and perturb signaling dynamics in living biological systems.

Author information

1
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 N. Wolfe St., Baltimore, MD 21205, USA.

Abstract

In this Commentary, we discuss two sets of genetically encoded molecular tools that have significantly enhanced our ability to observe and manipulate complex biochemical processes in their native context and that have been essential in deepening our molecular understanding of how intracellular signaling networks function. In particular, genetically encoded biosensors are widely used to directly visualize signaling events in living cells, and we highlight several examples of basic biosensor designs that have enabled researchers to capture the spatial and temporal dynamics of numerous signaling molecules, including second messengers and signaling enzymes, with remarkable detail. Similarly, we discuss a number of genetically encoded biochemical perturbation techniques that are being used to manipulate the activity of various signaling molecules with far greater spatial and temporal selectivity than can be achieved using standard pharmacological or genetic techniques, focusing specifically on examples of chemically driven and light-inducible perturbation strategies. We then describe recent efforts to combine these diverse and powerful molecular tools into a unified platform that can be used to elucidate the molecular details of biological processes that may potentially extend well beyond the realm of signal transduction.

KEYWORDS:

Biosensors; FRET; Live cell imaging; Signaling; Targeted perturbations

PMID:
24634506
PMCID:
PMC3953811
DOI:
10.1242/jcs.099994
[Indexed for MEDLINE]
Free PMC Article
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