Abstract

The post-transcriptional sequence modification of transcripts through RNA editing is an important mechanism for regulating protein function and is associated with human disease phenotypes. The identification of RNA editing or RNA-DNA difference (RDD) sites is a fundamental step in the study of RNA editing. However, a substantial number of false-positive RDD sites have been identified recently. A major challenge in identifying RDD sites is to distinguish between the true RNA editing sites and the false positives. Furthermore, determining the location of condition-specific RDD sites and elucidating their functional roles will help toward understanding various biological phenomena that are mediated by RNA editing. The present study developed RNA-sequence comparison and annotation for RNA editing (RCARE) for searching, annotating, and visualizing RDD sites using thousands of previously known editing sites, which can be used for comparative analyses between multiple samples. RCARE also provides evidence for improving the reliability of identified RDD sites. RCARE is a web-based comparison, annotation, and visualization tool, which provides rich biological annotations and useful summary plots. The developers of previous tools that identify or annotate RNA-editing sites seldom mention the reliability of their respective tools. In order to address the issue, RCARE utilizes a number of scientific publications and databases to find specific documentations respective to a particular RNA-editing site, which generates evidence levels to convey the reliability of RCARE. Sequence-based alignment files can be converted into VCF files using a Python script and uploaded to the RCARE server for further analysis. RCARE is available for free at http://www.snubi.org/software/rcare/.

Highlights

  • RNA editing is the post-transcriptional alteration of a single nucleotidesequence in primary messenger RNA or non-coding RNA including microRNA transcripts [1]

  • The most widespread type of RNA editing in mammals is A-to-I editing, which is mediated by the adenosine deaminases acting on RNA (ADAR) enzyme [2,3,4]

  • This study presented a new tool for RNA sequencing (RNA-Seq) comparison and annotation for RNA editing (RCARE), which determines condition-dependent RNA editing sites, provides rich systematic annotations and the evidence level of each RNA editing site

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Summary

Introduction

RNA editing is the post-transcriptional alteration of a single nucleotidesequence in primary messenger RNA (mRNA) or non-coding RNA (ncRNA) including microRNA (miRNA) transcripts [1]. ExpEdit increases reliability by using more reliable data compared toother prediction tools, but is not able to identify novel RNA editing sites It provides a user-friendly web interface for uploading raw RNA-Seq data such as FASTQ, SAM (sequence alignment map), and BAM (binary alignment map) files, and for exploring RNA editing. This study presented a new tool for RNA-Seq comparison and annotation for RNA editing (RCARE), which determines condition-dependent RNA editing sites, provides rich systematic annotations and the evidence level of each RNA editing site It identifies novel RNA editing sites in the same/individual DNA sequence and RNA-Seq variant call format (VCF) data, and delivers ‘executive summary’ plots for the annotation and comparison of multiple samples with RNA-Seq data through a userfriendly web interface. RCARE is freely available at http://www.snubi.org/ software/rcare/

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