Delivery of cationic polymer-siRNA nanoparticles for gene therapies in neural regeneration

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• 作者：Yanran Liang^a ; Zhonglin Liu^a ; ^{zhonglinliu@126.com} ; Xintao Shuai^b ; Weiwei Wang^b ; Jun Liu^a ; Wei Bi^c ; Chuanming Wang^a ; Xiuna Jing^a ; Yunyun Liu^a ; Enxiang Tao^a ; ^{taoenxiang@yahoo.com.cn}
• 关键词：PEG-PEI ; polyethylene glycol-polyethyleneimine ; NSCs ; neural stem cells ; NgR ; Nogo receptor ; siRNA ; small interfering RNA ; scr ; scrambled ; Lipo 2000 ; Lipofectamine 2000 ; GAPDH ; glyceraldehyde-3-phosphate dehydrogenase
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2012
• 期刊代码：159_0006291x
• 类别：bio
• 出版时间：18 May, 2012
• 卷：421
• 期：4
• 页码：690-695
• 文件大小：470 K

摘要

The therapeutic applications of neural stem cells (NSCs) have potential to promote recovery in many obstinate diseases in central nervous system. Regulation of certain gene expressions using siRNA may have significant influence on the fate of NSC. To achieve the optimum gene silencing effect of siRNA, non-viral vector polyethylene glycol-polyethyleneimine (PEG-PEI) was investigated in the delivery of siRNA to NSCs. The characteristics of PEG-PEI/siRNA polyplexes were detected by scanning electron microscopy (SEM). The effects of nanoparticles on cell viability were measured via CCK-8 assay. In addition, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry, and real-time PCR and Western Blot were employed to detect the gene inhibition effect of siRNA delivered by PEG-PEI. The SEM micrographs showed that PEG-PEI could condense siRNA to form diffuse and spherical nanoparticles. The cytotoxicity of PEG-PEI/siRNA nanocomplexes (N/P = 15) was significantly lower when compared with that of Lipofectamine 2000/siRNA (P < 0.05). Moreover, the highest transfection efficiency of PEG-PEI/siRNA nanoparticles was obtained at an N/P ratio of 15, which was better than that achieved in the transfection using Lipofectamine 2000 (P < 0.05). Finally, the gene knockdown effect of PEG-PEI/siRNA nanoparticles was verified at the levels of mRNA and protein. These results suggest that PEG-PEI may potentially be used as a siRNA delivery vector for neural regeneration therapy.