Identification of circRNAs for miRNA targets by Argonaute2 RNA immunoprecipitation and luciferase screening assays

Abstract

Circular RNAs from back-spliced exons (circRNAs) represent a novel class of widespread and endogenous RNAs in eukaryotes. circRNAs may bind to microRNAs (miRNAs) and inhibit the activity of miRNAs. Alternatively, miRNAs could also directly target circRNAs and regulate the expression of circRNAs. Here we describe the Argonaute2 (AGO2) RNA immunoprecipitation (RIP) and luciferase screening assays to identify the interaction between circRNAs and miRNAs. The AGO2 RIP assay evaluates the potential of the interaction between circRNAs and miRNAs. The luciferase screening assay investigates the targeting miRNAs and the detail binding sites for a specific circRNA.