Life under the Microscope: Single-Molecule Fluorescence Highlights the RNA World

The emergence of single-molecule (SM) fluorescence techniques has opened up a vast new toolbox for exploring the molecular basis of life. The ability to monitor individual biomolecules in real time enables complex, dynamic folding pathways to be interrogated without the averaging effect of ensemble...

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Bibliographic Details
Published in:Chemical reviews 2018-04, Vol.118 (8), p.4120-4155
Main Authors: Ray, Sujay, Widom, Julia R, Walter, Nils G
Format: Article
Language:English
Subjects:
Biological properties
Biology
Biomolecules
Catalysis
CRISPR-Cas Systems
Fluorescence
Fluorescence Resonance Energy Transfer
Folding
Gene Editing
Landscape
Microscopy, Fluorescence - methods
Molecules
Nucleic Acid Conformation
Protein Biosynthesis
Proteins
Reverse Transcription
Ribonucleic acid
Riboswitch
RNA
RNA - chemistry
RNA - genetics
RNA - physiology
RNA Interference
RNA Splicing
RNA-Binding Proteins - chemistry
RNA-protein interactions
Spectrometry, Fluorescence - methods
surveys
Telomerase - metabolism
Transcription, Genetic
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Summary:The emergence of single-molecule (SM) fluorescence techniques has opened up a vast new toolbox for exploring the molecular basis of life. The ability to monitor individual biomolecules in real time enables complex, dynamic folding pathways to be interrogated without the averaging effect of ensemble measurements. In parallel, modern biology has been revolutionized by our emerging understanding of the many functions of RNA. In this comprehensive review, we survey SM fluorescence approaches and discuss how the application of these tools to RNA and RNA-containing macromolecular complexes in vitro has yielded significant insights into the underlying biology. Topics covered include the three-dimensional folding landscapes of a plethora of isolated RNA molecules, their assembly and interactions in RNA-protein complexes, and the relation of these properties to their biological functions. In all of these examples, the use of SM fluorescence methods has revealed critical information beyond the reach of ensemble averages.
ISSN:0009-2665
1520-6890
1520-6890
DOI:10.1021/acs.chemrev.7b00519