Characterization of genetically encoded FRET biosensors for Rho-family GTPases

Sara K. Donnelly, Veronika Miskolci, Alice M. Garrastegui, Dianne Cox, Louis Hodgson

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages87-106
Number of pages20
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1821
ISSN (Print)1064-3745

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Keywords

  • Biosensors
  • FRET
  • Fluorometry
  • Ratiometric analysis
  • Rho GTPases
  • Validation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Donnelly, S. K., Miskolci, V., Garrastegui, A. M., Cox, D., & Hodgson, L. (2018). Characterization of genetically encoded FRET biosensors for Rho-family GTPases. In Methods in Molecular Biology (pp. 87-106). (Methods in Molecular Biology; Vol. 1821). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8612-5_7