Single-Molecule FRET X

Mike Filius; Raman van Wee; Chirlmin Joo

doi:10.1007/978-1-0716-3377-9_10

Single-Molecule FRET X

Mike Filius
, Raman van Wee
, Chirlmin Joo

Department of Physics

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

1 Scopus citations

Abstract

Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon that has been repurposed as a biophysical tool to measure nanometer distances. With FRET by DNA eXchange, or FRET X, many points of interest (POIs) in a single object can be probed, overcoming a major limitation of conventional single-molecule FRET. In FRET X, short fluorescently labeled DNA imager strands specifically and transiently bind their complementary docking strands on a target molecule, such that at most a single FRET pair is formed at each point in time and multiple POIs on a single molecule can be readily probed. Here, we describe the sample preparation, image acquisition, and data analysis for structural analysis of DNA nanostructures with FRET X.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	203-213
Number of pages	11
DOIs	https://doi.org/10.1007/978-1-0716-3377-9_10
State	Published - 2024

Publication series

Name	Methods in Molecular Biology
Volume	2694
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.

Keywords

DNA nanotechnology
FRET X
Single-molecule FRET
Single-molecule multiplexing
Structural biology

Access to Document

10.1007/978-1-0716-3377-9_10

Cite this

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keywords = "DNA nanotechnology, FRET X, Single-molecule FRET, Single-molecule multiplexing, Structural biology",

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AU - van Wee, Raman

AU - Joo, Chirlmin

N1 - Publisher Copyright: © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.

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AB - Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon that has been repurposed as a biophysical tool to measure nanometer distances. With FRET by DNA eXchange, or FRET X, many points of interest (POIs) in a single object can be probed, overcoming a major limitation of conventional single-molecule FRET. In FRET X, short fluorescently labeled DNA imager strands specifically and transiently bind their complementary docking strands on a target molecule, such that at most a single FRET pair is formed at each point in time and multiple POIs on a single molecule can be readily probed. Here, we describe the sample preparation, image acquisition, and data analysis for structural analysis of DNA nanostructures with FRET X.

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KW - Single-molecule multiplexing

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Single-Molecule FRET X

Abstract

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Keywords

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