Analyzing DNA Origami Nanostructure Assembly by Dynamic Light Scattering and Nanoparticle Tracking Analysis.

Area of Science:

• Biotechnology and Nanotechnology
• Molecular Biology and Self-Assembly

Background:

• Nucleic acid self-assembly enables precise fabrication of multi-dimensional nanostructures.
• These nanostructures have potential applications in biocatalysis, smart materials, molecular diagnostics, and therapeutics.

Purpose of the Study:

• To investigate DNA origami nanostructures using dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA).
• To compare the resolution and sensitivity of DLS and NTA for characterizing DNA nanostructures.
• To develop and validate a fluorescent NTA method for enhanced sensitivity.

Main Methods:

• Dynamic Light Scattering (DLS) and Nanoparticle Tracking Analysis (NTA) were employed.
• A novel fluorescent NTA method was developed using an intercalation dye.
• Analysis included 1D, 2D, and 3D DNA origami structures, as well as dynamic DNA nanocages.

Main Results:

• NTA provided higher resolution size measurements (smaller FWHM) compared to DLS for DNA nanostructures.
• The fluorescent NTA method demonstrated enhanced sensitivity for analyzing various DNA origami structures.
• NTA successfully characterized dynamic conformational switches in DNA nanocages.

Conclusions:

• NTA is a superior technique for high-resolution characterization of DNA nanostructures.
• Fluorescent NTA enhances sensitivity, crucial for quality control and mechanistic studies.
• Quantitative size distribution data from NTA provides critical insights into DNA nanomaterial applications.