Fig. 5

Methods to explore the possible mechanisms of circRNAs. a AGO2 CLIP-seq to predict the communication between miRNA and targeted circRNA. b, c circRNA-RBP interactions is mainly through RNA pull-down assay (b) or RNA immunoprecipitation (RIP) for experimental analysis (c). b In the RNA pull-down assay, a biotin-labeled probe recognized the BSJ of circRNA and then captured by biotin coupled magnetic beads. Finally, mass spectrum (MS) and western blot analysis to determinate the circRNA binding proteins. c In RIP-seq assay, the candidate RBP was first binds to the magnetic beads via antibody, and then the interacted circRNAs were analyzed by sequencing and RT-PCR. d circR-loops are identified by immunoprecipitation with the R loop-specific S9.6 antibody or catalytically inactive human RNase H1. Discovery the circR-loops in DRIP-seq data. circR-loops regulate diversity types of biological process, including transcriptional pausing, DNA damage, and double-strand DNA breaks (DSBs)