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

Mohammad Q. Hassan, Jonathan A.R. Gordon, Jane B. Lian, Andre J. Van Wijnen, Janet L. Stein, Gary S. Stein

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)817-822
Number of pages6
JournalJournal of cellular biochemistry
Volume110
Issue number4
DOIs
StatePublished - Jul 1 2010

Keywords

  • HLX1
  • RISC
  • RNP-IP
  • miR-27A
  • miR-Let-7/98

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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