DNA Origami Assembly Inside Liposomes via Nanopore‐Mediated DNA Transportation

Assembly of DNA origami nanostructures inside giant unilamellar vesicles (GUVs) is demonstrated by supplying approximately 200 unique staple strands from the external environment. Optimization of the lipid composition (DOPC : POPG : Cholesterol = 3:1:6) enables GUVs to withstand annealing for over 12 hours. Scaffold DNA is pre-encapsulated within the vesicles, and staple strands are transported through the pore-forming membrane protein streptolysin O. Subsequent addition of CaCl₂ induces pore closure, retaining scaffold and staple DNA strands inside GUVs during annealing. Agarose gel electrophoresis and atomic force microscopy confirm the formation of correctly assembled DNA origami nanostructures inside GUVs. This “bottle-ship” strategy opens possibilities for dynamically updating or installing new functions within compartments, beyond the simple delivery of energy or signal molecules.