title: 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.
authors.affiliation: Center for Integrated Protein Science Munich (CIPSM), Laboratory for RNA Biology, Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Stoehr
authors.forename: Gunter
authors.initials: G
authors.lastname: Meister
mesh_terms: D060565:Argonaute Proteins; D001681:Biological Assay; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D007553:Isotope Labeling; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D034743:RNA-Induced Silencing Complex / Q000378:metabolism*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D034743:RNA-Induced Silencing Complex; D012313:RNA