Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity

Luca Bordes; Sergei Chavez-Abiega; Joachim Goedhart

doi:10.1007/978-1-0716-1221-7_11

Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity

Luca Bordes
, Sergei Chavez-Abiega
, Joachim Goedhart^*

^*Corresponding author for this work

AIMMS

Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic › peer-review

Abstract

A wealth of assays for screening GPCR activity have been developed. Biosensors that employ Förster Resonance Energy transfer (FRET) are specific and enable dynamic measurements. Moreover, FRET biosensors are ideally suited for the analysis of single living cells. The FRET biosensors described in this manuscript are entirely genetically encoded by plasmids. Here, protocols for employing FRET-based biosensors to detect G protein activity upon GPCR activation are reported. The protocols include details on the isolation of plasmids, transfection, generation of stable cell lines with the FRET biosensors, FRET ratio imaging, and data analysis.

Original language	English
Title of host publication	G Protein-Coupled Receptor Screening Assays
Subtitle of host publication	Methods and Protocols
Editors	Sofia Aires M. Martins, Duarte Miguel F. Prazeres
Publisher	Humana Press Inc
Pages	159-178
Number of pages	20
Edition	2nd
ISBN (Electronic)	9781071612217
ISBN (Print)	9781071612200
DOIs	https://doi.org/10.1007/978-1-0716-1221-7_11
Publication status	Published - 2021

Publication series

Name	Methods in Molecular Biology
Volume	2268
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
S.C.-A. was funded by Marie Sklodowska-Curie Actions of the European Union?s Horizon 2020 program as part of the CaSR Biomedicine Network.

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Funding

S.C.-A. was funded by Marie Sklodowska-Curie Actions of the European Union?s Horizon 2020 program as part of the CaSR Biomedicine Network.

Funders
Horizon 2020 Framework Programme
HORIZON EUROPE Marie Sklodowska-Curie Actions

Keywords

Biosensor
FRET
GPCR
Image analysis
Ratio-imaging

Access to Document

10.1007/978-1-0716-1221-7_11

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Bordes, L., Chavez-Abiega, S., & Goedhart, J. (2021). Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity. In S. A. M. Martins, & D. M. F. Prazeres (Eds.), G Protein-Coupled Receptor Screening Assays: Methods and Protocols (2nd ed., pp. 159-178). (Methods in Molecular Biology; Vol. 2268). Humana Press Inc. https://doi.org/10.1007/978-1-0716-1221-7_11

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Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity. / Bordes, Luca; Chavez-Abiega, Sergei; Goedhart, Joachim.
G Protein-Coupled Receptor Screening Assays: Methods and Protocols. ed. / Sofia Aires M. Martins; Duarte Miguel F. Prazeres. 2nd. ed. Humana Press Inc, 2021. p. 159-178 (Methods in Molecular Biology; Vol. 2268).

Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic › peer-review

TY - CHAP

T1 - Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity

AU - Bordes, Luca

AU - Chavez-Abiega, Sergei

AU - Goedhart, Joachim

N1 - Funding Information: S.C.-A. was funded by Marie Sklodowska-Curie Actions of the European Union?s Horizon 2020 program as part of the CaSR Biomedicine Network. Publisher Copyright: © 2021, Springer Science+Business Media, LLC, part of Springer Nature. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021

Y1 - 2021

N2 - A wealth of assays for screening GPCR activity have been developed. Biosensors that employ Förster Resonance Energy transfer (FRET) are specific and enable dynamic measurements. Moreover, FRET biosensors are ideally suited for the analysis of single living cells. The FRET biosensors described in this manuscript are entirely genetically encoded by plasmids. Here, protocols for employing FRET-based biosensors to detect G protein activity upon GPCR activation are reported. The protocols include details on the isolation of plasmids, transfection, generation of stable cell lines with the FRET biosensors, FRET ratio imaging, and data analysis.

AB - A wealth of assays for screening GPCR activity have been developed. Biosensors that employ Förster Resonance Energy transfer (FRET) are specific and enable dynamic measurements. Moreover, FRET biosensors are ideally suited for the analysis of single living cells. The FRET biosensors described in this manuscript are entirely genetically encoded by plasmids. Here, protocols for employing FRET-based biosensors to detect G protein activity upon GPCR activation are reported. The protocols include details on the isolation of plasmids, transfection, generation of stable cell lines with the FRET biosensors, FRET ratio imaging, and data analysis.

KW - Biosensor

KW - FRET

KW - GPCR

KW - Image analysis

KW - Ratio-imaging

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DO - 10.1007/978-1-0716-1221-7_11

M3 - Chapter

C2 - 34085268

AN - SCOPUS:85107484550

SN - 9781071612200

T3 - Methods in Molecular Biology

SP - 159

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BT - G Protein-Coupled Receptor Screening Assays

A2 - Martins, Sofia Aires M.

A2 - Prazeres, Duarte Miguel F.

PB - Humana Press Inc

ER -

Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity

Abstract

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