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Characterization of Genetically Encoded FRET Biosensors for Rho-Family GTPases.

ABSTRACT: Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions to include additional/different regulators and inhibitors, as appropriate for the specific protein of interest.

SUBMITTER: Donnelly SK

PROVIDER: S-EPMC6104821 | biostudies-other | 2018

REPOSITORIES: biostudies-other

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Characterization of Genetically Encoded FRET Biosensors for Rho-Family GTPases.

Donnelly Sara K SK Miskolci Veronika V Garrastegui Alice M AM Cox Dianne D Hodgson Louis L

Methods in molecular biology (Clifton, N.J.) 20180101

Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions t ...[more]

PMID: 30062407

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