Title
Ribonucleoprotein immunoprecipitation (RNP-IP): a direct in vivo analysis of microRNA-targets
UMMS Affiliation
Department of Cell Biology and Cancer Center
Date
6-22-2010
Document Type
Article
Subjects
Immunoprecipitation; Ribonucleoproteins; MicroRNAs
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.
Rights and Permissions
Citation: J Cell Biochem. 2010 Jul 1;110(4):817-22. Link to article on publisher's site
Related Resources
PubMed ID
20564179
Journal Title
Journal of cellular biochemistry