Quantifying Local Molecular Tension Using Intercalated DNA Fluorescence

Nano Letters
Graeme A KingGijs J L Wuite

Abstract

The ability to measure mechanics and forces in biological nanostructures, such as DNA, proteins and cells, is of great importance as a means to analyze biomolecular systems. However, current force detection methods often require specialized instrumentation. Here, we present a novel and versatile method to quantify tension in molecular systems locally and in real time, using intercalated DNA fluorescence. This approach can report forces over a range of at least ∼0.5-65 pN with a resolution of 1-3 pN, using commercially available intercalating dyes and a general-purpose fluorescence microscope. We demonstrate that the method can be easily implemented to report double-stranded (ds)DNA tension in any single-molecule assay that is compatible with fluorescence microscopy. This is particularly useful for multiplexed techniques, where measuring applied force in parallel is technically challenging. Moreover, tension measurements based on local dye binding offer the unique opportunity to determine how an applied force is distributed locally within biomolecular structures. Exploiting this, we apply our method to quantify the position-dependent force profile along the length of flow-stretched DNA and reveal that stretched and entwined DNA ...Continue Reading

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Citations

Jan 29, 2019·Frontiers in Physiology·Laura TeodoriMaria C Albertini
May 9, 2021·Nucleic Acids Research·Dian SpakmanGraeme A King
May 25, 2021·ACS Sensors·Chandrashekhar U MuradeGeorge T Shubeita
Aug 13, 2021·Science Advances·Nicholas A W BellJustin E Molloy
Sep 11, 2021·The Journal of Physical Chemistry Letters·Natalie A PacePeter M Goodwin

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Methods Mentioned

BETA
Fluorescence
fluorescence microscopy
optical tweezers
fluorescence imaging

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