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circRNAs are known to be evolutionarily conserved and highly abundant, but few have been functionally validated. circRNAs are generated by linkage between a downstream 3'splice site and an upstream 5'splice site in a process known as backsplicing. In the first study, Pamudurti et al. generated ribosome footprinting (RFP) data from the head of Drosophila melanogaster, which they scanned for reads corresponding to circRNAs with predicted open reading frames (ORF) across the backspliced junction to identify those circRNAs bound by ribosomes (ribo-circRNAs).

The authors then transfected D. melanogaster S2 cells with contructs containing minigenes expressing some of the identified ribo-circRNAs: circMbl (one of multiple circRNAs generated from the muscleblind (mbl) locus), circCdi or circPde8. The minigenes were shown to produce proteins of expected size, and additional experiments confirmed that a …