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Genome repeat masking pipeline

This is a local pipeline to repeat-mask genomes before feeding them to https://github.com/nf-core/pairgenomealign. The rationale is that it is better when all soft masks have been produced by the same pipeline…

What it does:

This pipeline takes genomes as inputs and soft-masks their repeats with the following software:

  • tantan, version 51. Tantan is our default choice from a long time because TRF used to be non-free.
  • WindowMasker, version 1.0.0 distributed with BLAST 2.17.0.
  • RepeatMasker version 4.1.9.

The input of repeatmasker can be any of:

  • RepeatModeler version 2.0.7 (default)
  • Dfam (optional)
  • A custom repeat library (optional)

RepeatMasker and RepeatModeler are run from the same bioconda package as the standard nf-core module.

The pipeline then merges the soft masks of the RepeatMasker runs, and then merges that with the tantan and WindowMasker runs.

Finally, the pipeline prepares a MultiQC report that shows the extent of masking for each tool.

Disclaimer

This is not an official pipeline. This pipeline uses code and infrastructure developed and maintained by the nf-core initative, and reused here under the MIT license.

The nf-core framework for community-curated bioinformatics pipelines.

Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.

Nat Biotechnol. 2020 Feb 13. doi: 10.1038/s41587-020-0439-x.

Usage

This pipeline does not work with conda.

First make a sample sheet with usual nf-core pipelines.

samplesheet.csv:

sample,fasta
query_1,path-to-query-genome-file-one.fasta.gz
query_2,path-to-query-genome-file-two.fasta.gz

Genome files are uncompressed because WindowMasker does not handle stdin input.

Then run the pipeline as usual:

nextflow run oist/LuscombeU_stlrepeatmask \
   -profile oist \
   --input samplesheet.csv \
   --outdir <OUTDIR>

Test the dev branch of the pipeline (adapt the -w option for your own case!

nextflow run oist/LuscombeU_stlrepeatmask \
   -profile oist,test \
   -w /flash/LuscombeU/`whoami`/cache/deletemeTest \
   --outdir results_test

Test the local checkout

nextflow run ./main.nf \
   -profile oist,test \
   -w /flash/LuscombeU/`whoami`/cache/deletemeTest \
   --outdir results_test

Options

  • Point --repeatlib to a FASTA file to have an extra RepeatMasker run using it as a library.
  • Set --taxon to a taxon name to have an extra RepeatMasker run using the -species option set to that taxon.
  • Point --dfam to a directory containing a famdb subdirectory with the FamDB files in HDF5 format (not gzipped).

Dfam

Most containers that provide RepeatMasker do not contain a full copy of Dfam, which is huge. However they sometimes have a stub, for instance under /usr/local/share/RepeatMasker/Libraries or /opt/RepeatMasker/Libraries. Interstingly, when using the --libdir option, Dfam has to be in a subfolder named famdb, although it is named FamDB in the download website.

Pipeline output

tantan, repeatmodeler, windowmasker, dfam (optional), extlib (optional), mergedmasks

  • Masked genome file (compressed with bgzip).
  • BED file representing the masked regions.
  • Summary statistics of the softmasked genome.

Only in repeatmodeler

  • De novo detected repeats (.fa, .log, .stk and BLAST database files .n*)

Resource usage

On a test run on haplotype-merged and diploid assemblies of Oikopleura dioica (2n = 60 Mbp):

  • CPU usage was ~50 % for most processes. RepeatModeller was allocated 24 cores and used ~10 on average.
  • Memory usage was less than 1 GB for all processes except RepeatModeller (~6 GB, max 8 GB).
  • All processes needed only 10 % of the allocated time, except for RepeatModeller, which took between 100 and 500 minutes.
  • On a couple of primate genomes, RepeatModeller managed to keep its 24 cores 60% busy for ~30 hours using 40 GB memory.

Future directions

  • It may be interesting to add TRF and ULTRA, and compare and combine their results to the ones of tantan.

Credits

This pipeline was originally written by Mahdi and then taken over by @charles-plessy.

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Mask genomic repeats and low-complexity regions for the Scrambling in the Tree of Life project

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