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J Cell Biochem. 2010 Jul 1;110(4):817-22. doi: 10.1002/jcb.22562.

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

Author information

1
Department of Cell Biology and Cancer Center, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.

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.

PMID:
20564179
PMCID:
PMC3329753
DOI:
10.1002/jcb.22562
[Indexed for MEDLINE]
Free PMC Article

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