Ribonucleoprotein immunoprecipitation (RNP-IP): a direct in vivo analysis of microRNA-targets
Department of Cell Biology and Cancer Center
Immunoprecipitation; Ribonucleoproteins; MicroRNAs
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.
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Citation: J Cell Biochem. 2010 Jul 1;110(4):817-22. Link to article on publisher's site
Journal of cellular biochemistry
Hassan, Mohammad Q.; Gordon, Jonathan A. R.; Lian, Jane B.; Van Wijnen, Andre J.; Stein, Janet L.; and Stein, Gary S., "Ribonucleoprotein immunoprecipitation (RNP-IP): a direct in vivo analysis of microRNA-targets" (2010). Cell and Developmental Biology Publications and Presentations. Paper 92.