470495 Role of SNP Characteristics on the Endocytosis and Intracellular Trafficking of siRNA
Our goal is to define nanoparticle characteristics that are essential for potent siRNA delivery. Using silica nanoparticles (SNPs) for their tunable synthesis, we are investigating delivery criteria in four main categories: sequence specific mRNA degradation (silencing), membrane translocation, endosomal escape, and association with RNAi proteins. Using SNPs, vehicle size, structure, charge, and surface functionalization can be varied to determine optimal vehicle design criteria in a variety of cell types.
The presentation will discuss our novel live cell assay that allows independent intracellular trafficking of delivery vehicle, antisense, and sense siRNA. Using confocal microscopy, we track vehicle-siRNA complexes during endocytosis (whether by Clathrin, Caveolin, ARF6, GRAF1, Flotillin, and Macropinocytosis), endosomal trafficking (Early, Late, Recycling, and Lysosome), trafficking to the endoplasmic reticulum or golgi, and exocytosis. Our results to date demonstrate that multiple vehicle characteristics affect siRNA delivery and demonstrate the critical role of intracellular trafficking in maximizing siRNA potency.