Extensive alternative splicing (AS) of precursor mRNAs (pre-mRNAs) in multicellular eukaryotes increases the protein-coding capacity of a genome and allows novel ways to regulate gene expression. In flowering plants, up to 48% of intron-containing genes exhibit AS. However, the full extent of AS in plants is not yet known, as only a few high-throughput RNA-Seq studies have been performed.  As the cost of obtaining RNA-Seq reads continues to fall, it is anticipated that huge amounts of plant sequence data will accumulate and help in obtaining a more complete picture of AS in plants. Although it is not an onerous task to obtain hundreds of millions of reads using high-throughput sequencing technologies, computational tools to accurately predict and visualize AS are still being developed and refined.

This review discusses the tools to predict and visualize transcriptome-wide AS in plants using short-reads and highlight their limitations. Comparative studies of AS events between plants and animals have revealed that there are major differences in the most prevalent types of AS events, suggesting that plants and animals differ in the way they recognize exons and introns. Extensive studies have been performed in animals to identify cis-elements involved in regulating AS, especially in exon skipping. However, few such studies have been carried out in plants. Here, the current state of research on splicing regulatory elements (SREs) is reviewed and emerging experimental and computational tools to identify cis-elements involved in regulation of AS in plants are discussed.

The availability of curated alternative splice forms in plants makes it possible to use computational tools to predict SREs involved in AS regulation, which can then be verified experimentally. Such studies will permit identification of plant-specific features involved in AS regulation and contribute to deciphering the splicing code in plants.

Tools for predicting isoforms, their expression, and alternative splicing from RNA-Seq data.

Method Task Input data
Trans-ABySS (Robertson et al., 2010) IP, IE De novo
Trinity (Grabherr et al., 2011) IP, IE De novo
Rnnotator (Martin et al., 2010) IP De novo
Scripture (Guttman et al., 2010) IP G
IsoLasso (Li et al., 2011) IP, IE G
NSMAP (Xia et al., 2011) IP, IE G
Cufflinks (Trapnell et al., 2010) IP, IE G, A
TAU (Filichkin et al., 2010) IP G,A
SpliceGrapher (Rogers et al., 2012) SG G, A
IsoEM (Nicolae et al., 2010) IE G, A
IsoformEX (Kim et al., 2011) IE G, A
SpliceTrap (Wu et al., 2011) IE G, A
NEUMA (Lee et al., 2011) IE G, A
Solas (Richard et al., 2010) IE G, A
rSeq (Jiang and Wong, 2009) IE G, A
RSEM (Li et al., 2010; Li and Dewey, 2011) IE De novo

 

The tools vary in the specific task they address; we distinguish between several tasks: isoform prediction (IP), isoform expression (IE) and splice graph prediction (SG). The tools also vary in the input data they require: de novo (no input required except for the RNA-Seq data), a reference genome (G) or annotated isoforms (A).

  • Reddy AS, Rogers MF, Richardson DN, Hamilton M, Ben-Hur A. (2012) Deciphering the plant splicing code: experimental and computational approaches for predicting alternative splicing and splicing regulatory elements. Front Plant Sci 3, 18. [article]

Incoming search terms:

  • splicing
  • alternative splicing
  • high throughput sequencing
  • paenms seq
  • paenms seq gob mx
  • PAENMS SEQ GOB
  • paenms seq go
  • how to test rnnotator
  • www paenms seq gop
  • alternative splicing plant rna-seq

Comments

Leave a Reply




  • Social Networking Pages

    Linkedin Group

  • Follow Me on Pinterest
  • RSS SEQanswers – RNA Sequencing

    • Identifying small RNA sequence within whole genome sequence May 21, 2013
      Hi all, I want to know if there are any useful bioinformatic tool to find small RNA sequence within a whole bacteria genome. Thank you in... […]
      Inma
    • standard of clean data May 21, 2013
      Hi all I recently got my prokaryotes RNA-seq data report back. the standard filter steps of the raw data set by our local sequencing center is as... […]
      Pengfei Liu
    • Problem with cummeRbund diffData() May 20, 2013
      Hi all, I'm running Tophat/cufflinks/cuffdiff for differential gene expression and analysis with cummeRbund (v 2.0.0). I'm having an issue with... […]
      Enrique Zudaire
    • How to increase rowsize in heatmap? May 16, 2013
      Hi, I am a complete newbie to all things cummeRbund and am currently fighting with generating readable heatmaps. When I use ... […]
      Mags
    • novoalign mapping May 15, 2013
      Hi, I want to use novoalign to map reads - allowing up to 15 mismatches for 100 bp paired-end reads I am new to novoalign(went through the... […]
      abh
    • Design of expt across multiple lanes May 15, 2013
      Hi, I am performing an RNA-seq experiment to look at differential expression. The design is as follows: 2 populations x 3 biological... […]
      jbono
  • RSS Biostar – RNA-Seq

    • What are the best practices for SNP identification in RNA seq transcriptome data
      I have 20 RICE RNA seq tranascriptome data hiseq 2000 platform paired end reads. I aligned fasta reads with BWA and remove PCR duplicates with PICARD. Later I call SNP with samtools using various parameters. I would like to clarify what parameters should I used while alinging to reference rice genome for looking SNP location 100 bp upstream and 250 bp downst […]
    • How do TopHat options -g , --supress-hits, and Bowtie options interplay?
      Hi, I am currently using TopHat2 to map RNA-seq runs. I think there have been some changes pertaining the -g option. Does anyone know how it works now? I used to think that setting -g would look for n alignments for a given read, report them [if top-scoring] and discard those reads that had more than g [top scoring] alignments. Now, the description sounds mo […]
    • What happened to -k in TopHat for multiple-mapping reads?
      Selecting -g n in tophat does not discard reads mapping more than n, but instead only reports n alignments for those out all all their TOP scoring alignments. I think there used to be an option -k that would allow one to discard reads that topped x alignments -- whatever happened to that? I only see -g in the tophat 2 manual, no reporting options like before […]
    • Does tophat use the library-type information for mapping, or just for the XS flag?
      When I specify library-type to TopHat, i.e., first-strand, second-strand, unstranded, TopHat appends a value + or - to the XS:A tag, which is useful for subsequent analyses, such as annotation. However, does this information influence the "mappability" of reads, or is this unaffected? My guess is that the information will be considered for mapping […]
    • Purpose of Y-shaped adapters in Illumina Sequencing?
      Hi all, Y adapters different sequences to be annealed to the 5' and 3' ends of each molecule in a library. The arms of the Y are unique, and the middle part, connected to the DNA fragment, is complementary. What are the advantages of this? My take of this over having fully-complementary adapters (ADAPTER1 - - - - - ADAPTER1) is that: -Upon primer a […]
    • Cell Type composition in a tissue based on gene marker expression
      I am not sure if the following would even make sense.... Tissues are composed of composite cell types, and often there are studies such as microarray/NGS where we perform a collective sampling of cells from these tissues. Information about the composition (say percentage of cell type) is not taken into consideration. In some case (such as brain/cancer), ther […]