LEMONS - A Tool for the Identification of Splice Junctions in Transcriptomes of Organisms Lacking Reference Genomes

Liron Levin, Dan Bar-Yaacov, Amos Bouskila, Michal Chorev, Liran Carmel, Dan Mishmar

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

RNA-seq is becoming a preferred tool for genomics studies of model and non-model organisms. However, DNA-based analysis of organisms lacking sequenced genomes cannot rely on RNA-seq data alone to isolate most genes of interest, as DNA codes both exons and introns. With this in mind, we designed a novel tool, LEMONS, that exploits the evolutionary conservation of both exon/intron boundary positions and splice junction recognition signals to produce high throughput splice-junction predictions in the absence of a reference genome. When tested on multiple annotated vertebrate mRNA data, LEMONS accurately identified 87% (average) of the splice-junctions. LEMONS was then applied to our updated Mediterranean chameleon transcriptome, which lacks a reference genome, and predicted a total of 90,820 exon-exon junctions. We experimentally verified these splice-junction predictions by amplifying and sequencing twenty randomly selected genes from chameleon DNA templates. Exons and introns were detected in 19 of 20 of the positions predicted by LEMONS. To the best of our knowledge, LEMONS is currently the only experimentally verified tool that can accurately predict splice-junctions in organisms that lack a reference genome.

Original languageEnglish
Article numbere0143329
Pages (from-to)e0143329
JournalPLoS ONE
Volume10
Issue number11
DOIs
StatePublished - 1 Nov 2015

Keywords

  • Computational Biology/methods
  • Exons
  • Genomics/methods
  • Introns
  • Nucleotide Motifs
  • RNA Splice Sites
  • RNA Splicing
  • RNA, Messenger/genetics
  • Reproducibility of Results
  • Software
  • Transcriptome

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