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

• Published in: Biochemical and biophysical research communications 2012-05, Vol.421 (4), p.690-695
• Main Authors: Liang, Yanran, Liu, Zhonglin, Shuai, Xintao, Wang, Weiwei, Liu, Jun, Bi, Wei, Wang, Chuanming, Jing, Xiuna, Liu, Yunyun, Tao, Enxiang
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Animals; CENTRAL NERVOUS SYSTEM; DISEASES; FLUORESCENCE; Gene Knockdown Techniques; GENE THERAPY; GENES; Genetic Therapy - methods; GPI-Linked Proteins - genetics; MESSENGER-RNA; Mice; Myelin Proteins - genetics; Nanoparticles - administration & dosage; Nanoparticles - chemistry; NANOSTRUCTURES; NERVE CELLS; Nerve Regeneration; Neural stem cell; Neurons - physiology; Nogo Receptor 1; Non-viral vector; PHOSPHATES; Polyethylene glycol-polyethyleneimine; POLYETHYLENE GLYCOLS; Polyethylene Glycols - chemistry; Polyethyleneimine - analogs & derivatives; Polyethyleneimine - chemistry; POLYMERASE CHAIN REACTION; RECEPTORS; Receptors, Cell Surface - genetics; RNA, Small Interfering - administration & dosage; RNA, Small Interfering - chemistry; siRNA; STEM CELLS; TOXICITY; Transfection
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2012.03.155

► Nogo receptor can inhibit growth of injured axons, thus affecting neural regeneration. ► The delivery of siRNA is crucial to inhibit NgR expression in NSCs. ► Non-viral vector PEG-PEI condensed siRNA targeting NgR into nanoscale particles. ► PEG-PEI/siRNA at N/P=15 displayed high transfection efficiency and low cytotoxicity. ► PEG-PEI has great potential in carrying siRNA to diminish the gene expression in NSCs. 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