Abstract

BackgroundAlternative splicing is an important cellular mechanism that can be analyzed by RNA sequencing. However, identification of splicing events in an automated fashion is error-prone. Thus, further validation is required to select reliable instances of alternative splicing events (ASEs). There are only few tools specifically designed for interactive inspection of ASEs and available visualization approaches can be significantly improved.ResultsHere, we present Manananggal, an application specifically designed for the identification of splicing events in next generation sequencing data. Manananggal includes a web application for visual inspection and a command line tool that allows for ASE detection. We compare the sashimi plots available in the IGV Viewer, the DEXSeq splicing plots and SpliceSeq to the Manananggal interface and discuss the advantages and drawbacks of these tools. We show that sashimi plots (such as those used by the IGV Viewer and SpliceSeq) offer a practical solution for simple ASEs, but also indicate short-comings for highly complex genes.ConclusionManananggal is an interactive web application that offers functions specifically tailored to the identification of alternative splicing events that other tools are lacking. The ability to select a subset of isoforms allows an easier interpretation of complex alternative splicing events. In contrast to SpliceSeq and the DEXSeq splicing plot, Manananggal does not obscure the gene structure by showing full transcript models that makes it easier to determine which isoforms are expressed and which are not.

Highlights

  • Alternative splicing is an important cellular mechanism that can be analyzed by RNA sequencing

  • Since the tool includes bodymap data, we decided to go for an alternative splicing event in PKIG between heart and brain that we will use for the comparison of another tool later

  • According to the GTEX portal and the tissue specific data stored in SpliceSeq, different isoforms of PKIG are expressed in brain and heart that use two different promoters

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Summary

Introduction

Alternative splicing is an important cellular mechanism that can be analyzed by RNA sequencing. Evidence has been gathered for the involvement of alternative splicing in neurological disorders (e.g. autism [2], Huntington’s disease [3], spinal muscular atrophy [4]), autoimmune diseases (e.g. multiple sclerosis [5], systemic lupus erythematosus [6], Kawasaki disease [7]) and tumorigenesis [8,9,10,11]. Understanding this aberrant splicing behavior could translate into a health benefit for patients

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