PACeR: a bioinformatic pipeline for the analysis of chimeric RNA-seq data

Abstract

ABSTRACTMicroRNAs (miRNAs) are small non-coding RNAs that function in post-transcriptional gene regulation through imperfect base pairing with mRNA targets which results in inhibition of translation and often destabilization of bound transcripts. Sequence-based algorithms historically used to predict miRNA targets face inherent challenges in reliably reflecting in vivo interactions. Recent strategies have directly profiled miRNA-target interactions by cross-linking and ligation of miRNAs to their targets within the RNA-induced silencing complex (RISC), followed by high throughput sequencing of the chimeric RNAs. Despite the strength of these direct chimeric miRNA:target profiling approaches, standardized pipelines for analyzing the resulting chimeric RNA sequencing data are not readily available. Here we present PACeR, a robust bioinformatic pipeline for the analysis of chimeric RNA sequencing data. PACeR consists of two parts, each of which are optimized for the distinctive characteristics of chimeric RNA sequencing reads: first, read processing and alignment and second, peak calling and motif analysis. We apply PACeR to chimeric RNA sequencing data generated in our lab as well as a published benchmark dataset. PACeR has minimal computational power requirements and contains extensive annotation to broaden accessibility for processing chimeric RNA sequencing data and enable insights to be gained about the targets of small non-coding RNAs in regulating diverse biological systems.