SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation

Abstract

cGMP is critical to a variety of cellular processes, but the available tools to interfere with endogenous cGMP lack cellular and subcellular specificity. We introduce SponGee, a genetically encoded chelator of this cyclic nucleotide that enables in vitro and in vivo manipulations in single cells and in biochemically defined subcellular compartments. SponGee buffers physiological changes in cGMP concentration in various model systems while not affecting cAMP signals. We provide proof-of-concept strategies by using this tool to highlight the role of cGMP signaling in vivo and in discrete subcellular domains. SponGee enables the investigation of local cGMP signals in vivo and paves the way for therapeutic strategies that prevent downstream signaling activation.

Keywords: (T)hPDE5(VV); FRET; PKG; axon guidance; cGMP buffer; genetically encoded; lipid grafts; neuronal migration; single cell pharmacology; subcellular compartment.