31st European Congress on Nanotechnology and Materials Engineering
Institute of Chemical Technology, India
Title: Eliciting in vitro delivery of siRNA & CRISPR using biodegradable surface modified Chitosan coated PLGA Nanoparticles
Biography: Ashu Srivastav
The development of safe and optimally efficient delivery system for siRNA and CRISPR/ Cas9 elements capable of achieving specific targeting for gene therapy is a major challenge in clinical therapeutics, therefore we developed biodegradable Chitosan modified PLGA nanoparticles that can efficiently deliver siRNA and Cas9/sgRNA to the cells. Owing to high positive charge and low toxicity of CS-PLGA NPs, they have high loading capacity as well as high transfection efficiency for therapeutic delivery. CS-PLGA NPs were prepared by using solvent-emulsion method and PF-68 was used as surfactant and siRNA and Cas9/sgRNA was loaded onto them. The physio-chemical properties such as particle size and Zeta potential were measured by DLS and their surface morphology was assessed using SEM and TEM. The cytotoxicity of NPs as delivery system was investigated on human cervical cancer HeLa cells using MTT assay which indicated that NPs had no potential toxic effects on cell viability. The binding efficiency was validated by Gel retardation Assay. Internalization studies for fluorescent Cy3 siRNA and FITC Cas9/sgRNA loaded with CS-PLGA were observed using confocal microscopy. In vitro silencing assay of Hela GFP stable cells was performed using formulated NPs loaded with Turbo-GFP siRNA and it confirmed the transfection efficiency of NPs. This study suggested that biodegradable cationic CS-PLGA nanocomplex possess great potential for efficient and safe therapeutic delivery and can be used as a promising delivery agent.