Abstract

BackgroundTranslational regulation is one important aspect of gene expression regulation. Dysregulation of translation results in abnormal cell physiology and leads to diseases. Ribosome profiling (RP), also called ribo-seq, is a powerful experimental technique to study translational regulation. It can capture a snapshot of translation by deep sequencing of ribosome-protected mRNA fragments. Many ribosome profiling data processing tools have been developed. However, almost all tools analyze ribosome profiling data at the gene level. Since different isoforms of a gene may produce different proteins with distinct biological functions, it is advantageous to analyze ribosome profiling data at the isoform level. To meet this need, previously we developed a pipeline to analyze 610 public human ribosome profiling data at the isoform level and constructed HRPDviewer database.ResultsTo allow other researchers to use our pipeline as well, here we implement our pipeline as an easy-to-use software tool called RPiso. Compared to Ribomap (a widely used tool which provides isoform-level ribosome profiling analyses), our RPiso (1) estimates isoform abundance more accurately, (2) supports analyses on more species, and (3) provides a web-based viewer for interactively visualizing ribosome profiling data on the selected mRNA isoforms.ConclusionsIn this study, we developed RPiso software tool (http://cosbi7.ee.ncku.edu.tw/RPiso/) to provide isoform-level ribosome profiling analyses. RPiso is very easy to install and execute. RPiso also provides a web-based viewer for interactively visualizing ribosome profiling data on the selected mRNA isoforms. We believe that RPiso is a useful tool for researchers to analyze and visualize their own ribosome profiling data at the isoform level.

Highlights

  • Translational regulation is one important aspect of gene expression regulation

  • Using RPiso to analyze ribo-seq data from a cell cycle study in Hela cell [19], we have shown in our HRPDviewer study [16] that the cyclin G1 mRNA isoform NM_004060 is constitutively translated throughout the cell cycle with peak levels at early G1 phase but the other cyclin G1 mRNA isoform NM_199246 is only translated in G1 phase, indicating that the two mRNA isoforms of the gene CCNG1 may be under different translational regulations in the cell cycle process

  • RSEM was originally designed for RNA-seq, we have shown in our HRPDviewer database paper [16] that RSEM can be used to handle read mapping uncertainty for Ribo-seq with high accuracy

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Summary

Results

To allow other researchers to use our pipeline as well, here we implement our pipeline as an easy-to-use software tool called RPiso. Compared to Ribomap (a widely used tool which provides isoform-level ribosome profiling analyses), our RPiso (1) esti‐ mates isoform abundance more accurately, (2) supports analyses on more species, and (3) provides a web-based viewer for interactively visualizing ribosome profiling data on the selected mRNA isoforms

Conclusions
Background
Results and discussion
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