新型双嵌段磷酸胆碱基共聚物作为siRNA基因运输载体研究

Study on phosphorylcholine diblock copolymers as siRNA gene carriers

将载体MPC30-DEA70(N)与siRNA(P)在N/P比值不同的条件下复合,用DNA凝胶电泳法,倒置显微镜,MTT法表征复合物溶液;同时分析胎牛血清(FBS)对细胞转染的影响。结果显示在无胎牛血清条件下siRNA能与载体MPC30—DEA70电性中和,N/P比值越大中和的越多,而有胎牛血清的条件下siRNA几乎不能与载体MPC30-DEA70电性中和;倒置显微镜观察到siRNA分子能进入细胞,转染效率随着N/P比值增大而增大;同时随着载体浓度增大,对细胞的增殖抑制性增强。以上结果显示,新型阳离子磷酸胆碱聚合物MPC30-DEA70可以有效负载和运输siRNA,是一种安全有效的非病毒类转基因载体。

MPC30—DEA70/siRNA complexes were formed at different N/P ratios（cationic/anionic） conditions.The complexes were studied using gel electrophoresis,FCM,and MTT assay.Results showed some polyplexes are neutralized in the absence of FBS,significantly with the increasing of N/P ratios.Confocal laser scanning microscopy found the transfected siRNA could enter into the cells,suggesting its effective nuclear localization and the transfection efficiency were significantly enhanced with increasing of N/P ratios.These results indicate that Novel cationic phosphorylcholine copolymer can load and transport siRNA,which may act as an efficient and safe non—viral gene transfer vector.