一种新颖的阳离子非病毒基因载体Chitosan-g-PEI-g-PEG-OH的性能研究

Evaluation of the Properties of a Novel Cationic Non-viral Gene Carrier Chitosan-g-PEI-g-PEG-OH

研究了新型阳离子聚合物Chitosan-g-PEI-g-PEG-OH的性能,重点考察其粒度,基因转染效率与细胞毒性,探讨了其作为基因载体的可能性.通过动态光散射仪（DSL）、透射电镜（TEM）观察了Chitosan-g-PEI-g-PEG-OH与DNA自组装形成的颗粒形态及粒径,Chitosan-g-PEI-g-PEG-OH可复合DNA形成粒径160～210 nm的纳米复合物,适合进入细胞.使用MTT比色法分析Chitosan-g-PEI-g-PEG-OH的毒性并与PEI,PEI-g-PEG-OH比较.选用增强型绿色荧光蛋白（EGFP）转染Hela细胞,应用流式细胞术检测转染效率.新型阳离子多聚物Chitosan-g-PEI-g-PEG-OH在提高基因转染效率的同时降低了其细胞毒性,有望成为基因转移的有效载体.

Chitosan-g-PEI-g-PEG-OH is a new cationic copolymer.The influencing factors of Chitosan-g-PEI-g-PEG-OH gene delivery efficiency were studied,and these properties demonstrated that this copolymer could be used as the gene carrier.The surface topography and sizes of self-assembly complexes of the Chitosan-g-PEI-g-PEG-OH and DNA were observed through DSL and TEM,and the main size of the Chitosan-g-PEI-g-PEG-OH/DNA complex was approximately 160～210 nm spherical particles.The particles with the size were suitable for entry into cells.Cytotoxicity of Chitosan-g-PEI-g-PEG-OH was evaluated in MTT method and compared with Cytotoxicity of PEI and PEI-g-PEG-OH.Chitosan-g-PEI-g-PEG-OH/pEGFP complexes were transfected into Hela,and the effects of the transfection conditions were evaluated with FACS.Chitosan-g-PEI-g-PEG-OH copolymer presented a lower cytotoxicity and higher transfection efficiency than PEI.The results show that the copolymer is quite promising as an efficient carrier for gene delivery.