title: Native gel analysis for RISC assembly.
abstract: Small-interfering RNAs (siRNAs) and microRNAs (miRNAs) regulate expression of their target mRNAs via the RNA-induced silencing complex (RISC). A core component of RISC is the Argonaute (Ago) protein, which dictates the RISC function. In Drosophila, miRNAs and siRNAs are generally loaded into Ago1-containing RISC (Ago1-RISC) and Ago2-containing RISC (Ago2-RISC), respectively. We developed a native agarose gel system to directly detect Ago1-RISC, Ago2-RISC, and their precursor complexes. Methods presented here will provide powerful tools to biochemically dissect the RISC assembly pathways.
authors.affiliation: Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kawamata
authors.forename: Yukihide
authors.initials: Y
authors.lastname: Tomari
mesh_terms: D000818:Animals; D060565:Argonaute Proteins; D004330:Drosophila / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D039642:Eukaryotic Initiation Factors / Q000378:metabolism; D007553:Isotope Labeling; D009154:Mutation / Q000235:genetics; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D020365:RNA Helicases / Q000235:genetics; D034743:RNA-Induced Silencing Complex / Q000378:metabolism*; D043244:Ribonuclease III / Q000235:genetics
chemical_list: C489654:AGO1 protein, Drosophila; D060565:Argonaute Proteins; D029721:Drosophila Proteins; D039642:Eukaryotic Initiation Factors; D034743:RNA-Induced Silencing Complex; D012313:RNA; C478069:DCR-2 protein, Drosophila; D043244:Ribonuclease III; D020365:RNA Helicases