Chitosan-graft-polyethylenimine as a gene carrier.
Chitosans have been proposed as biocompatible alternative cationic polymers that are suitable for non-viral delivery. However, the transfection efficiency of chitosan-DNA nanoparticles is still very low. To improve transfection efficiency, we prepared chitosan-graft-polyethylenimine (CHI-g-PEI) copolymer by an imine reaction between periodate-oxidized chitosan and polyethylenimine (PEI). The molecular weight and composition of the CHI-g-PEI copolymer were characterized, using multi-angle laser scattering (GPC-MALS) and (1)H nuclear magnetic resonance ((1)H NMR), respectively. The copolymer was complexed with plasmid DNA (pDNA) in various copolymer/DNA (N/P) charge ratios, and the complex was characterized. CHI-g-PEI showed good DNA binding ability and high protection of DNA from nuclease attack. Also, with an increase in charge ratio, the sizes of the CHI-g-PEI/DNA complex showed a tendency to decrease, whereas the zeta potential of the complex showed an increase. The CHI-g-PEI copolymer had low cytotoxicity, compared to PEI 25K from cytotoxicity assays. At high N/P ratios, the CHI-g-PEI/DNA complex showed higher transfection efficiency than PEI 25K in HeLa, 293T and HepG2 cell lines. Our results indicate that the CHI-g-PEI copolymer has potential as a gene carrier in vitro.
Jiang
Hu-Lin
HL
School of Agricultural Biotechnology, Seoul National University, Seoul, South Korea.
Kim
You-Kyoung
YK
Arote
Rohidas
R
Nah
Jae-Woon
JW
Cho
Myung-Haing
MH
Choi
Yun-Jaie
YJ
Akaike
Toshihiro
T
Cho
Chong-Su
CS
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000138:chemical synthesis / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D018942:Macrolides / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D010316:Particle Size; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry*; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry; D012995:Solubility; D055672:Static Electricity; D013499:Surface Properties; D014162:Transfection / Q000379:methods*
D004791:Enzyme Inhibitors; D018942:Macrolides; D011108:Polymers; C040929:bafilomycin A1; D011094:Polyethyleneimine; D048271:Chitosan; D008156:Luciferases; D003850:Deoxyribonuclease I