Development of probes for cellular functions using fluorescent proteins and fluorescence resonance energy transfer

Many genetically encoded probes that employ fluorescent proteins and fluorescence resonance energy transfer (FRET) have been developed to better understand the spatiotemporal regulation of various cellular processes. The different types of FRET and measurement techniques necessitate characterization...

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Bibliographic Details
Published inAnnual review of biochemistry Vol. 80; p. 357
Main Author Miyawaki, Atsushi
Format Journal Article
LanguageEnglish
Published United States 07.07.2011
Subjects
Animals
Cell Physiological Phenomena
Fluorescence Resonance Energy Transfer - methods
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
Green Fluorescent Proteins - chemistry
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Microscopy, Fluorescence - methods
Models, Molecular
Protein Conformation
Proteins
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Summary:Many genetically encoded probes that employ fluorescent proteins and fluorescence resonance energy transfer (FRET) have been developed to better understand the spatiotemporal regulation of various cellular processes. The different types of FRET and measurement techniques necessitate characterization of their specific features. Here I provide theoretical and practical comparisons of bimolecular and unimolecular FRET constructs, intensity-based and lifetime-based FRET measurements, FRET imaging using live- and fixed-cell samples, green fluorescent protein-based and chemical fluorophore-based FRET, and FRET efficiency and indices. The potential benefits and limitations of a variety of features in the technologies using fluorescent proteins and FRET are discussed.
ISSN:1545-4509
DOI:10.1146/annurev-biochem-072909-094736