Identifying molecular targets of endogenous ADAR3 in gliolastoma cells

Through immunoprecipitation of epitope-tagged ADAR3 protein, several ADAR3-bound RNAs have been reported till date. However, exogenous expression of proteins are usually associated with altered gene expression profiles and thus, can influence binding profiles of RNA-binding proteins (RBPs) being studied. Here, we used affinity-purified antibody to pull down ADAR3 endogenously expressed in U373 glioblastoma cells and identified transcripts significantly enriched in the ADAR3 immunoprecipitants over control. To serve as control, antisera isolated from rabbit prior to immunization with ADAR3 antigen was used to perform the IP. Overall design: RNA Immunoprecipitation-Sequencing (RIP-Seq) for endogenously-expressed ADAR3 protein in U373 glioblastoma cells.