Plasmid DNA complexation with phosphorylcholine diblock copolymers and its effect on cell transfection.
We examined a series of novel cationic MPC-based (2-methacryloyloxyethyl phosphorylcholine) copolymers as vectors for gene delivery, with emphasis on the assessment of the effects of the charge ratio (administered via pH variation) on the complex (polyplex) formation and the subsequent transfection efficiency. A combination of electrophoresis, dynamic light scattering, and small angle neutron scattering was used to characterize the structure and charge distribution of the polyplexes formed between the copolymer and the luciferase plasmid DNA. Polymers with larger hydrophobic side chains had lower p K a values and tended to aggregate more strongly. For a given copolymer, electrostatic interaction was the main driving force for the formation of the nanopolyplexes. When the cationic copolymers were in excess, the majority of the polyplexes formed was neutral, and only a small faction of them carried net positive charges. Polyplexes formed under excess copolymer protected the DNA from restriction enzyme digestion. As the copolymers were weak polyelectrolytes, the pH had a distinct effect on the structure and charge distribution of the polyplexes formed. Below the p K a, the copolymers were found to bind with the plasmid DNA in the form of unimers, while above the p K a, the copolymers self-aggregated and complexed with DNA in the form of micelles. It was subsequently found that unimer/DNA polyplexes were far more effective in the transfection of HEK293 cells than micellar DNA polyplexes. The results thus revealed that different hydrophobicities of the side chains in the copolymer series led to different nanostructuring and charge characteristics, which had a consequential effect on the transfection efficiency. This study provided useful insight into the molecular processes underlying polyplex formation and demonstrated a strong link between structural and physical properties of polyplexes and cell transfection efficiency.
Zhao
Xiubo
X
Biological Physics Group, School of Physics and Astronomy, The University of Manchester, Schuster Building, Manchester M13 9PL, United Kingdom.
Zhang
ZhuoQi
Z
Pan
Fang
F
Waigh
Thomas A
TA
Lu
Jian R
JR
D002460:Cell Line; D004247:DNA / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010767:Phosphorylcholine / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D012996:Solutions; D014162:Transfection
D011108:Polymers; D012996:Solutions; D010767:Phosphorylcholine; D004247:DNA