Polymer based gene silencing: In vitro delivery of SiRNA

Abstract

Gene silencing may be achieved by harnessing the RNA interference mechanism to effect down-regulation of protein expression. The therapeutic use of siRNA is dependent on its delivery to the intracellular space. This chapter describes the delivery of siRNA by N-(2-ethylamino)-6-O-glycolchitosan (EAGC). EAGC is a chitosan-based polymer, which binds to siRNA to form nanoparticles (NPs). The steps necessary to determine the delivery capacity of a polymer are presented in this chapter using EAGC as an example. The steps include: the transfection of cells with EAGC-siRNA polyplexes and protein detection by a Western Blotting assay.