FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs

Bryce T. Bajar; Xinmeng Guan; Amy Lam; Michael Z. Lin; Ryohei Yasuda; Tal Laviv; Jun Chu

doi:10.1007/978-1-0716-2035-9_2

FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs

Bryce T. Bajar, Xinmeng Guan, Amy Lam, Michael Z. Lin, Ryohei Yasuda, Tal Laviv, Jun Chu^*

^*Corresponding author for this work

Physiology Pharmacology

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

1 Scopus citations

Abstract

With the development of fluorescent proteins (FPs) and advanced optical microscopy techniques, Förster or fluorescence resonance energy transfer (FRET) has become a powerful tool for real-time noninvasive visualization of a variety of biological processes, including kinase activities, with high spatiotemporal resolution in living cells and organisms. FRET can be detected in appropriately configured microscopes as changes in fluorescence intensity, lifetime, and anisotropy. Here, we describe the preparation of samples expressing FP-based FRET sensors for RhoA kinase, intensity- and lifetime-based FRET imaging, and postimaging data analysis.

Original language	English
Title of host publication	Cell Polarity Signaling
Editors	Chenbei Chang, Jianbo Wang
Publisher	Humana Press Inc.
Pages	31-43
Number of pages	13
ISBN (Electronic)	978-1-0716-2035-9
ISBN (Print)	978-1-0716-2034-2
DOIs	https://doi.org/10.1007/978-1-0716-2035-9_2
State	Published - 2022

Publication series

Name	Methods in Molecular Biology
Volume	2438
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Funding

Funders	Funder number
Max Planck Florida Institute for Neuroscience
NSFC of Guangdong Province Shenz-hen Science and Technology Innovation Committee	KQJSCX20170331161420421, JCYJ20170818164040422, JCYJ20170818163925063
NSFC of Guangdong Province Shenzhen Science and Technology Innovation Committee
National Institutes of Health	DP1DP1NS096787, R01MH080047, F30EY029952, T32GM008042
National Institute of Neurological Disorders and Stroke	R35NS116804
Human Frontier Science Program
National Natural Science Foundation of China	31670872, 21874145, 81927803
National Key Research and Development Program of China	2017YFA0700403

Keywords

FLIM-FRET
FRET
Fluorescence lifetime
Fluorescent protein
Rho GTPase
RhoA
Sensitized emission FRET

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10.1007/978-1-0716-2035-9_2

Cite this

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abstract = "With the development of fluorescent proteins (FPs) and advanced optical microscopy techniques, F{\"o}rster or fluorescence resonance energy transfer (FRET) has become a powerful tool for real-time noninvasive visualization of a variety of biological processes, including kinase activities, with high spatiotemporal resolution in living cells and organisms. FRET can be detected in appropriately configured microscopes as changes in fluorescence intensity, lifetime, and anisotropy. Here, we describe the preparation of samples expressing FP-based FRET sensors for RhoA kinase, intensity- and lifetime-based FRET imaging, and postimaging data analysis.",

keywords = "FLIM-FRET, FRET, Fluorescence lifetime, Fluorescent protein, Rho GTPase, RhoA, Sensitized emission FRET",

author = "Bajar, {Bryce T.} and Xinmeng Guan and Amy Lam and Lin, {Michael Z.} and Ryohei Yasuda and Tal Laviv and Jun Chu",

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doi = "10.1007/978-1-0716-2035-9_2",

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AU - Yasuda, Ryohei

AU - Laviv, Tal

AU - Chu, Jun

PY - 2022

Y1 - 2022

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AB - With the development of fluorescent proteins (FPs) and advanced optical microscopy techniques, Förster or fluorescence resonance energy transfer (FRET) has become a powerful tool for real-time noninvasive visualization of a variety of biological processes, including kinase activities, with high spatiotemporal resolution in living cells and organisms. FRET can be detected in appropriately configured microscopes as changes in fluorescence intensity, lifetime, and anisotropy. Here, we describe the preparation of samples expressing FP-based FRET sensors for RhoA kinase, intensity- and lifetime-based FRET imaging, and postimaging data analysis.

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FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs

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Keywords

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