DNA Origami Seesaws as Comparative Binding Assay.

Area of Science:

• Biophysics
• Nanotechnology
• Molecular Biology

Background:

• Traditional force spectroscopy methods in biology require invasive tethers.
• These tethers connect molecules of interest to external measurement devices like beads or cantilever tips.
• This limits the scope and accuracy of biological force measurements.

Purpose of the Study:

• To develop a novel, non-invasive force spectroscopy technique using DNA origami.
• To demonstrate its utility in comparative binding assays.
• To enable in situ molecular interaction analysis without physical connections.

Main Methods:

• Designed a DNA origami seesaw-like nanostructure with a freely moving lever.
• Utilized binding partners on each side of the structure to induce distinct conformations.
• Performed model experiments with competing DNA hybridization reactions.

Main Results:

• The DNA origami structure shifted to conformations dictated by binding partner strength.
• Stronger binding interactions resulted in a pronounced conformational shift.
• Demonstrated tuneable reference DNA duplexes for comparative measurements.

Conclusions:

• The DNA origami prototype offers an in situ readout for molecular binding.
• This non-invasive approach overcomes limitations of traditional force spectroscopy.
• The device is suitable for measuring ligand interactions in comparative assays.