Encapsulation of DNA by cationic diblock copolymer vesicles

Abstract

Encapsulation of dsDNA fragments (contour length 54 nm) by the cationic diblock copolymer poly(butadiene-b-N-methyl 4-vinyl pyridinium) [PBd-b-P4VPQ] has been studied with phase contrast, polarized light, and fluorescence microscopy, as well as scanning electron microscopy. Encapsulation was achieved with a single emulsion technique. For this purpose, an aqueous DNA solution is emulsified in an organic solvent (toluene) and stabilized by the amphiphilic diblock copolymer. The PBd block forms an interfacial brush, whereas the cationic P4VPQ block complexes with DNA. A subsequent change of the quality of the organic solvent results in a collapse of the PBd brush and the formation of a capsule. Inside the capsules, the DNA is compacted as shown by the appearance of birefringent textures under crossed polarizers and the increase in fluorescence intensity of labeled DNA. The capsules can also be dispersed in aqueous medium to form vesicles, provided they are stabilized with an osmotic agent (polyethylene glycol) in the external phase. It is shown that the DNA is released from the vesicles once the osmotic pressure drops below 105 N/m2 or if the ionic strength of the supporting medium exceeds 0.1 M. The method has also proven to be efficient to encapsulate pUC18 plasmid in sub-micron sized vesicles and the general applicability of the method has been demonstrated by the preparation of the charge inverse system: cationic poly(ethylene imine) encapsulated by the anionic diblock poly(styrene-b-acrylic acid).