Single-Molecule FRET X

Book chapter (2023)

Authors

M. Filius BN/Chirlmin Joo Lab - , Kavli institute of nanoscience Delft

R.G. van Wee Kavli institute of nanoscience Delft, BN/Chirlmin Joo Lab -

C. Joo Kavli institute of nanoscience Delft, BN/Chirlmin Joo Lab -

Research Group

BN/Chirlmin Joo Lab () (TU Delft)

DOI: https://doi.org/10.1007/978-1-0716-3377-9_10

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

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Published Date

2023

Language

English

Reuse Rights

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Faculty

Applied Sciences

Department

BN/Bionanoscience

Research Group

BN/Chirlmin Joo Lab

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.

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