title: Native gel analysis for mammalian microRNPs assembled from pre-microRNAs.
abstract: MicroRNAs (miRNAs) are an important class of small RNAs that regulate gene expression posttranscriptionally through the microRNP (miRNP)/RNA-induced silencing complex (RISC). The core component of miRNPs is an Argonuate protein that directly binds to a miRNA. In mammals, most miRNPs are assembled through the miRNA loading complex (miRLC), which is composed of Dicer, Ago, and TRBP. miRLC processes miRNA precursors (pre-miRNAs) into miRNA duplexes and loads miRNA duplexes to Ago. Here, we describe a native gel analysis system for detecting miRNPs assembled with pre-miRNAs from mammalian lysates that ectopically express Ago2. The methods presented here provide a powerful tool for further dissecting miRNP assembly pathways in mammals.
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104-6100, USA.
authors.forename: Xuhang
authors.initials: X
authors.lastname: Liu
authors.forename: Zissimos
authors.initials: Z
authors.lastname: Mourelatos
mesh_terms: D000818:Animals; D053487:DEAD-box RNA Helicases / Q000235:genetics* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D055786:Gene Knockout Techniques / Q000379:methods*; D008322:Mammals; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D034743:RNA-Induced Silencing Complex / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D043244:Ribonuclease III / Q000235:genetics* / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D034743:RNA-Induced Silencing Complex; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases