Molecular force spectroscopy with a DNA origami-based nanoscopic force clamp

Forces in biological systems are typically investigated at the single-molecule level with atomic force microscopy or optical and magnetic tweezers, but these techniques suffer from limited data throughput and their requirement for a physical connection to the macroscopic world. We introduce a self-a...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2016-10, Vol.354 (6310), p.305-307
Main Authors: Nickels, Philipp C., Wünsch, Bettina, Holzmeister, Phil, Bae, Wooli, Kneer, Luisa M., Grohmann, Dina, Tinnefeld, Philip, Liedl, Tim
Format: Article
Language:English
Subjects:
Bending
Biomolecules
Clamps
Deoxyribonucleic acid
DNA
DNA, Cruciform - chemistry
DNA, Cruciform - ultrastructure
DNA, Single-Stranded - chemistry
DNA, Single-Stranded - ultrastructure
Fluorescence Resonance Energy Transfer - methods
Gene Expression Regulation
Molecular biology
Molecular structure
Nanostructure
Nanotechnology
Nanotechnology - methods
Promoter Regions, Genetic
Protein Binding
Sensor arrays
Single Molecule Imaging - methods
Spectrum analysis
Springs
Stress, Mechanical
TATA-Box Binding Protein - chemistry
TATA-Box Binding Protein - ultrastructure
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Summary:Forces in biological systems are typically investigated at the single-molecule level with atomic force microscopy or optical and magnetic tweezers, but these techniques suffer from limited data throughput and their requirement for a physical connection to the macroscopic world. We introduce a self-assembled nanoscopic force clamp built from DNA that operates autonomously and allows massive parallelization. Single-stranded DNA sections of an origami structure acted as entropic springs and exerted controlled tension in the low piconewton range on a molecular system, whose conformational transitions were monitored by single-molecule Förster resonance energy transfer. We used the conformer switching of a Holliday junction as a benchmark and studied the TATA-binding protein-induced bending of a DNA duplex under tension. The observed suppression of bending above 10 piconewtons provides further evidence of mechanosensitivity in gene regulation.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.aah5974