Synthesis and characterization of PEG-b-(PG-g-PEI) for gene delivery
- VernacularTitle:可降解非病毒基因载体PEG-b-（PG-g-PEI）的合成与表征
- Author:
Ning HE
1
;
Hechun SUN
;
Huanxi XU
;
Xiaoman DONG
;
Zhangzhang SHAO
Author Information
1. 安徽中医药大学药学院
- Keywords:
non-viral gene carrier;
polyethylenimine;
transfection
- From:
Journal of Southern Medical University
2013;(11):1643-1647
- CountryChina
- Language:Chinese
-
Abstract:
Objective To synthesize a biodegradable non-viral gene carrier with a high transfection efficiency and a low cytotoxicity. Methods Poly(ethylene glycol)- block- (poly(L- glutamic acid)- graft- polyethylenimine) was prepared via ammonolysis of poly(ethylene glycol)-block-poly (γ-benzyl L-glutamate) with the low-molecular-mass polyethylenimine (600 Da). The synthesized copolymer was characterized by 1H nuclear magnetic resonance spectroscopy and gel permeation chromatography. The polyplex micelle from PEG-b-(PG-g-PEI) and plasmid DNA (pDNA) was studied using dynamic light scattering, zeta-potential measurements, and gel retardation assay. The in vitro cytotoxicity and transfection efficiency of PEG-b-(PG-g-PEI) were tested by MTT assay and luciferase assay in HEK 293T cells using PEI (25 kDa) as the control. Results PEG-b-(PG-g-PEI) could efficiently condense DNA into nanosized particles with positive surface charges when the N/P ratio of polymer and DNA was above 5:1. The zeta potential of the polyplexes was about 25 mV, and the particle size was 120 nm at a N/P ratio of 10. The cell toxicity and gene transfection evaluations showed a lower cytotoxicity and a higher gene transfection efficiency of the copolymer than PEI 25000 in HEK 293T cells. Conclusions The polymer can be used as a potential non-viral gene carrier for gene therapy.
