In Vitro RISC Cleavage Assay

Abstract

RNA interference (RNAi) is a widely used tool for the analysis of gene expression. In this process, short-interfering RNAs (siRNAs) guide the RNA-induced silencing complex (RISC) to complementary target RNA molecules, which are sequence-specifically cleaved by the RISC. In vitro cleavage assays have proved to be powerful tools for the characterization of the RNAi pathway in many different organisms. Therefore, this chapter provides a detailed protocol for in vitro RISC assays.