Comparative analysis of circular RNA enrichment methods

The circRNAs sequencing results vary due to the different enrichment methods and their performance is needed to systematic comparison. This study investigated the effects of different circRNA enrichment methods on sequencing results, including abundance and species of circRNAs, as well as the sensitivity and precision. This experiment was carried out by following four common circRNA enrichment methods: including ribosomal RNA depletion (rRNA(–)), polyadenylation and poly (A(+)) RNA depletion followed by RNase R treatment (polyA+RNase R), rRNA(–)+polyA+RNase R and polyA+RNase R+ rRNA(–). The results showed that polyA+RNase R+ rRNA (–) enrichment method obtained more circRNA number, higher sensitivity and abundance among them; polyA+RNase R method obtained higher precision. The linear RNAs can be thoroughly removed in all enrichment methods except rRNA depletion method. Overall, our results helps researchers to quickly selection a circRNA enrichment of suitable for own study among many enrichment methods, and it provides a benchmark framework for future improvements circRNA enrichment methods. [Figure: see text]