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.
DOI of Published Version
J Cell Biochem. 2010 Jul 1;110(4):817-22. Link to article on publisher's site
Journal of cellular biochemistry
Hassan MQ, Gordon JA, Lian JB, Van Wijnen AJ, Stein JL, Stein GS. (2010). Ribonucleoprotein immunoprecipitation (RNP-IP): a direct in vivo analysis of microRNA-targets. Cell and Developmental Biology Publications. https://doi.org/10.1002/jcb.22562. Retrieved from https://escholarship.umassmed.edu/cellbiology_pp/92