Ribonucleoprotein immunoprecipitation (RNP-IP): A direct in vivo analysis of microRNA-targets

Academic Article

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. © 2010 Wiley-Liss, Inc.
  • Authors

    Published In

    Digital Object Identifier (doi)

    Author List

  • Hassan MQ; Gordon JAR; Lian JB; Van Wijnen AJ; Stein JL; Stein GS
  • Start Page

  • 817
  • End Page

  • 822
  • Volume

  • 110
  • Issue

  • 4