Probing the nanoparticle-AGO2 interaction for enhanced gene knockdown

RNAi is emerging as a promising technology for treatment of various diseases due to its ability to silence specific target genes. To date, a number of nanoparticle based formulations have been reported for the delivery of small interfering RNA (siRNA), with continuous modifications in the nanoparticle design for enhancing their efficiency. While majority of the design aspects are focused on avoiding or overcoming endosomal entrapment, limited studies are available that address the role of interaction of nanoparticles with the RNA induced silencing complex (RISC) machinery, which is a crucial aspect deciding the outcome. Here, we systematically probed the effect of steric hindrance of nanoparticles on RISC interaction, by modulating two parameters, nanoparticle size and hardness. An assay was developed for quantifying the extent of RISC interaction of different nanoparticles in vitro, which was then correlated with their gene knockdown efficiency. The results suggest that the soft and small nanoparticles were most efficacious in knocking down polo-like-kinase 1 (PLK1) siRNA, a gene overexpressed in a variety of cancer types.