Circular RNAs (circRNAs) are involved in various biological processes and disease pathogenesis. However, only a small number of functional circRNAs have been identified among hundreds of thousands of circRNA species, partly because most current methods are based on circular junction counts and overlook the fact that circRNA is formed from the host gene by back-splicing (BS). To distinguish the expression difference originated from BS or the host gene, Peking University researchers have developed DEBKS, a software program to streamline the discovery of differential BS events between two rRNA-depleted RNA sequencing (RNA-seq) sample groups. By applying real and simulated data and employing RT-qPCR for validation, the researchers demonstrate that DEBKS is efficient and accurate in detecting circRNAs with differential BS events between paired and unpaired sample groups.
A. The workflow of DEBKS consists of four modules: merge, count, anno, and dec. B. The schematic diagram of back-splicing. Red reads represent CJCs. Green reads represent LJCs. CJC, circular junction count; LJC, linear junction count; L, RNA-seq read length minus the minimum overhang length; e, circRNA length.
Availability – DEBKS is available at https://github.com/yangence/DEBKS as open-source software.