Characterization of genetically encoded FRET biosensors for Rho-family GTPases

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

doi:10.1007/978-1-4939-8612-5_7

Characterization of genetically encoded FRET biosensors for Rho-family GTPases

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

Anatomy & Structural Biology

Research output: Chapter in Book/Report/Conference proceeding › Chapter

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 language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	87-106
Number of pages	20
DOIs	https://doi.org/10.1007/978-1-4939-8612-5_7
State	Published - Jan 1 2018

Publication series

Name	Methods in Molecular Biology
Volume	1821
ISSN (Print)	1064-3745

Keywords

Biosensors
FRET
Fluorometry
Ratiometric analysis
Rho GTPases
Validation

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-4939-8612-5_7

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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

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