Cellular Uptake of DNA Origami

Area of Science:

• Biotechnology
• Nanotechnology
• Cell Biology

Background:

• Cells naturally internalize extracellular materials.
• Efficient intracellular delivery of DNA origami nanoparticles (DONs) is challenging due to harsh cellular environments.
• Standard cell culture media can degrade unprotected DONs.

Purpose of the Study:

• To provide comprehensive methods for achieving and analyzing cellular uptake of DNA origami.
• To outline protocols for preparing DNA origami for intracellular delivery.
• To detail post-uptake analysis techniques for DONs.

Main Methods:

• Sterile handling and endotoxin-free scaffolds for DNA origami folding.
• Stabilization strategies to protect DONs in cell culture media.
• Protocols for confocal microscopy and flow cytometry analysis of cellular uptake.
• Methods for assessing intracellular fate and retention of DONs.

Main Results:

• Successful cellular uptake of DNA origami is achievable through optimized protocols.
• Stabilization is crucial for DONs to survive extracellular and intracellular environments.
• Specific preparation and analysis methods correlate with successful uptake quantification.

Conclusions:

• This chapter provides a detailed guide to enhance the efficiency of DNA origami cellular uptake.
• The described methods facilitate reliable analysis of DONs within cells.
• Optimized protocols are essential for advancing DNA origami applications in nanomedicine and cell biology.