logo

HOMER

Software for motif discovery and next-gen sequencing analysis

RNA Motif Analysis

HOMER was not originally designed with RNA in mind, but it can be used to successfully analyze data for RNA motifs.  By RNA motifs, we mean short sequence elements in RNA sequences akin to DNA motifs, not structural elements such as hairpins and stuff like that.  For example, HOMER can be used to successfully determine miRNA seeds in sets of co-regulated mRNAs, or RNA binding elements in CLIP-Seq data.

The "-rna" option can be used with findMotifs.pl and findMotifsGenome.pl, resulting in + strand only motif searching and motif display/matching with "U" instead of "T".  HOMER does not contain a list of well known "RNA motifs" yet, so no "known motif" analysis is performed.  If using FASTA files, please use "T" (normal DNA encoding) in the input files for now.

Analyzing Co-regulated Gene Lists for RNA motifs

HOMER contains preconfigured PROMOTER sets comprised of RefSeq mRNA sequences, or only the 5' and 3' UTRs.  These are useful for analyzing gene lists for motifs in their mRNAs.  To run the analysis, us findMotifs.pl with a mRNA PROMOTER set, and options for RNA motifs will be automatically set.

findMotifs.pl mir1-downregulated.genes.txt human-mRNA MotifOutput/ -rna -len 8

You don't actually need to specify -rna for this case since with the use of "human-mRNA" it's understood.  Anyway, the output will look something like this:

mir1 example motifs

For now, HOMER will try to match the results to the human list of miRNA seeds (from miRBase):

matches to mir1 motifs


In this case, the motif matches the miR-1 consensus seed (which is shared by miR-206 and miR-613).

There are two RNA specific options for findMotifs.pl in rna mode:
-min <#> : minimum length of mRNA to consider (basically removes extremely short mRNA sequences from the analysis)
-max <#> : maximum length of mRNA to consider (removes really long RNAs from the analysis)

Analyzing CLIP-Seq for RNA motifs

HOMER can analyze strand-specific genomic regions for motifs, such as the regions that would be defined from CLIP-Seq.  To do this, just run findMotifsGenome.pl using the "-rna" flag (make sure your regions are strand specific!!).  For now, HOMER just uses the same random genomic background used for ChIP-Seq motif finding.  You could imagine that a better RNA motif finding background would be RNA, i.e. strand specific exon/intron sequences.  You'd be right, but managing this with respect to the different experiments (i.e. intronic binding vs. mRNA binding vs. non-coding RNA binding) is tricky and for now left up to the user (you can specify your own strand specific background with "-bg <peak/BED file>").  Trying this with FOX CLIP-Seq data:

findMotifsGenome.pl fox2.clip.bed hg17 MotifOutput -rna

This will give the following results (which resembles a UGCAUG FOX motif):

FOX RNA binding motif
Can't figure something out? Questions, comments, concerns, or other feedback:
cbenner@ucsd.edu