title: Ribonucleoprotein immunoprecipitation (RNP-IP): a direct in vivo analysis of microRNA-targets.
abstract: We have developed a novel Ribonucleoprotein Immunoprecipitation (RNP-IP) method to isolate miR-RISC complexes, associated microRNAs and target mRNAs. Our method characterizes mRNAs present in immunoprecipitates containing miR-RISC complexes that were obtained using GW182 and AGO2 antibodies. MicroRNA bound transcripts were reverse transcribed and amplified using seed sequence and 3'UTR derived primers. This flexible IP-based assay is a straightforward method to identify miRs participating in gene regulation and their cognate mRNAs in real time.
authors.affiliation: Department of Cell Biology and Cancer Center, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
authors.forename: Mohammad Q
authors.initials: MQ
authors.lastname: Hassan
authors.forename: Jonathan A R
authors.initials: JA
authors.lastname: Gordon
authors.forename: Jane B
authors.initials: JB
authors.lastname: Lian
authors.forename: Andre J
authors.initials: AJ
authors.lastname: van Wijnen
authors.forename: Janet L
authors.initials: JL
authors.lastname: Stein
authors.forename: Gary S
authors.initials: GS
authors.lastname: Stein
mesh_terms: D020413:3' Untranslated Regions; D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D047468:Immunoprecipitation; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D017931:DNA Primers; D035683:MicroRNAs; D012261:Ribonucleoproteins