Cellular Uptake of DNA Origami.

This chapter discusses the methods involved in achieving and analyzing cellular uptake of DNA origami. While cells naturally internalize substances from their surroundings, more than a simple addition of DNA origami in the surrounding cell medium is necessary to ensure DNA origami particles successfully enter the intracellular environment. Starting with the folding of the DNA, careful handling of sterile buffers and tools is essential, as well as the use of an endotoxin free scaffold. We explain how DNA origami needs a certain form of stabilization or protection to survive the degrading low-salt and high-nuclease environment of common cell culture media. Depending on the preferred method of post-uptake analysis (confocal), microscopy, or flow cytometry, we elaborate on the full protocols and crucial steps to prepare cell uptake experiments. Finally, notes are added on the intracellular fate (see Notes 14 and 15), and cellular retention of DNA origami (see Note 16) is discussed.