A Microsphere-Supported Lipid Bilayer Platform for DNA Reactions on a Fluid Surface

Area of Science:

• Biotechnology
• Nanotechnology
• Molecular Biology

Background:

• DNA nanotechnologies like toehold-mediated strand displacement (TMSD) and DNAzymes are crucial for molecular logic and dynamic systems.
• These reactions are typically performed in solution, limiting their integration into surface-based platforms.
• Developing fluid, stable surface platforms is essential for advancing DNA nanotechnology.

Purpose of the Study:

• To create a versatile microsphere-supported lipid bilayer (μSLB) platform for implementing DNA nanotechnologies on a fluid surface.
• To demonstrate the utility of this platform for TMSD and DNAzyme reactions.
• To enable multiplexed detection and improve reaction kinetics for DNA-based molecular systems.

Main Methods:

• Functionalizing microspheres with supported lipid bilayers (μSLBs) with membrane-bound nucleic acids.
• Optimizing reaction conditions to minimize DNA-phospholipid interactions and enhance bilayer stability.
• Tuning lipid composition and incorporating fluorescent lipids for flow cytometry readout.

Main Results:

• Successfully implemented both TMSD and DNAzyme reactions on the μSLB platform.
• Developed a multiplexed TMSD Dengue serotyping assay detecting all 16 sequence patterns with a 3 nM limit of detection.
• Demonstrated enhanced diffusion and kinetics for DNAzyme reactions on the fluid lipid surface.

Conclusions:

• The μSLB system provides a robust and versatile platform for surface-based DNA nanotechnologies.
• This platform facilitates the expansion of TMSD and DNAzyme reactions for molecular logic and advanced DNA nanotechnology.
• The system's multiplexability and fluid surface properties offer potential for compartmentalization and improved reaction kinetics.