DROPA: DRIP-seq optimized peak annotator

BackgroundR-loops are three-stranded nucleic acid structures that usually form during transcription and that may lead to gene regulation or genome instability. DRIP (DNA:RNA Immunoprecipitation)-seq techniques are widely used to map R-loops genome-wide providing insights into R-loop biology. However, annotation of DRIP-seq peaks to genes can be a tricky step, due to the lack of strand information when using the common basic DRIP technique.ResultsHere, we introduce DRIP-seq Optimized Peak Annotator (DROPA), a new tool for gene annotation of R-loop peaks based on gene expression information. DROPA allows a full customization of annotation options, ranging from the choice of reference datasets to gene feature definitions. DROPA allows to assign R-loop peaks to the DNA template strand in gene body with a false positive rate of less than 7%. A comparison of DROPA performance with three widely used annotation tools show that it identifies less false positive annotations than the others.ConclusionsDROPA is a fully customizable peak-annotation tool optimized for co-transcriptional DRIP-seq peaks, which allows a finest gene annotation based on gene expression information. Its output can easily be integrated into pipelines to perform downstream analyses, while useful and informative summary plots and statistical enrichment tests can be produced.

[1]  F. Chédin Nascent Connections: R-Loops and Chromatin Patterning. , 2016, Trends in genetics : TIG.

[2]  Xiaoqin Xu,et al.  Prevalent, Dynamic, and Conserved R-Loop Structures Associate with Specific Epigenomic Signatures in Mammals. , 2016, Molecular cell.

[3]  G. Capranico,et al.  DNA damage and genome instability by G-quadruplex ligands are mediated by R loops in human cancer cells , 2018, Proceedings of the National Academy of Sciences.

[4]  Clifford A. Meyer,et al.  Model-based Analysis of ChIP-Seq (MACS) , 2008, Genome Biology.

[5]  K. Cimprich,et al.  R-Loops as Cellular Regulators and Genomic Threats. , 2019, Molecular cell.

[6]  C. Glass,et al.  Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. , 2010, Molecular cell.

[7]  Aaron R. Quinlan,et al.  Bioinformatics Applications Note Genome Analysis Bedtools: a Flexible Suite of Utilities for Comparing Genomic Features , 2022 .

[8]  Andrés Aguilera,et al.  R loops: new modulators of genome dynamics and function , 2015, Nature Reviews Genetics.

[9]  I. Korf,et al.  R-loop formation is a distinctive characteristic of unmethylated human CpG island promoters. , 2012, Molecular cell.

[10]  Rasiah Loganantharaj,et al.  PAVIS: a tool for Peak Annotation and Visualization , 2013, Bioinform..

[11]  T. Swigut,et al.  Transcription-Replication Conflict Orientation Modulates R-Loop Levels and Activates Distinct DNA Damage Responses , 2017, Cell.

[12]  Hilde van der Togt,et al.  Publisher's Note , 2003, J. Netw. Comput. Appl..

[13]  Robert Gentleman,et al.  Software for Computing and Annotating Genomic Ranges , 2013, PLoS Comput. Biol..

[14]  F. Chédin,et al.  DNA Topoisomerase I differentially modulates R-loops across the human genome , 2018, Genome Biology.

[15]  Carsten Kuenne,et al.  UROPA: a tool for Universal RObust Peak Annotation , 2017, Scientific Reports.