nf-core/circrna

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Introduction

nf-core/circrna is a bioinformatics best-practice analysis pipeline for Quantification, miRNA target prediction and differential expression analysis of circular RNAs.

The pipeline is built using Nextflow, a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The Nextflow DSL2 implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from nf-core/modules in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community!

On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources.The results obtained from the full-sized test can be viewed on the nf-core website.

Pipeline summary

• Raw read QC (FastQC)
• Adapter trimming (Trim Galore!)
• MultiQC report MultiQC
• circRNA quantification
• CIRIquant
• STAR 2-Pass mode
  • CIRCexplorer2
  • circRNA finder
  • DCC
• find circ
• MapSplice
• Segemehl
• circRNA annotation
• Export mature spliced length as FASTA file
• Annotate parent gene, underlying transcripts.
• circRNA count matrix
• miRNA target prediction
  • miRanda
  • TargetScan
  • Filter results, miRNAs must be called by both tools
• Differential expression analysis DESeq2
• Circular - Linear ratio tests ‘CircTest’

Quick Start

1. Install Nextflow (>=22.10.1)

2. Install any of Docker, Singularity (you can follow this tutorial), Podman, Shifter or Charliecloud for full pipeline reproducibility (you can use Conda both to install Nextflow itself and also to manage software within pipelines. Please only use it within pipelines as a last resort; see docs).

3. Download the pipeline and test it on a minimal dataset with a single command:

   nextflow run nf-core/circrna -profile test,YOURPROFILE --outdir <OUTDIR>

   nextflow run nf-core/circrna -profile test,YOURPROFILE --outdir <OUTDIR>

   Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (YOURPROFILE in the example command above). You can chain multiple config profiles in a comma-separated string.

   • The pipeline comes with config profiles called docker, singularity, podman, shifter, charliecloud and conda which instruct the pipeline to use the named tool for software management. For example, -profile test,docker.
   • Please check nf-core/configs to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use -profile <institute> in your command. This will enable either docker or singularity and set the appropriate execution settings for your local compute environment.
   • If you are using singularity, please use the nf-core download command to download images first, before running the pipeline. Setting the NXF_SINGULARITY_CACHEDIR or singularity.cacheDir Nextflow options enables you to store and re-use the images from a central location for future pipeline runs.
   • If you are using conda, it is highly recommended to use the NXF_CONDA_CACHEDIR or conda.cacheDir settings to store the environments in a central location for future pipeline runs.

4. Start running your own analysis!

   nextflow run nf-core/circrna --input samplesheet.csv --outdir <OUTDIR> --genome GRCh37 -profile <docker/singularity/podman/shifter/charliecloud/conda/institute> --tool 'ciriquant' --module 'circrna_discovery,mirna_prediction,differential_expression' --bsj_reads 2

   nextflow run nf-core/circrna --input samplesheet.csv --outdir <OUTDIR> --genome GRCh37 -profile <docker/singularity/podman/shifter/charliecloud/conda/institute> --tool 'ciriquant' --module 'circrna_discovery,mirna_prediction,differential_expression' --bsj_reads 2

Documentation

The nf-core/circrna pipeline comes with documentation about the pipeline usage, parameters and output.

nextflow run nf-core/circrna \
   -profile <docker/singularity/.../institute> \
   --input samplesheet.csv \
   --outdir <OUTDIR>

nextflow run nf-core/circrna \
   -profile <docker/singularity/.../institute> \
   --input samplesheet.csv \
   --outdir <OUTDIR>

For more details and further functionality, please refer to the usage documentation and the parameter documentation.

Pipeline output

To see the results of an example test run with a full size dataset refer to the results tab on the nf-core website pipeline page. For more details about the output files and reports, please refer to the output documentation.

Credits

nf-core/circrna was originally written by Barry Digby.

We thank the following people for their extensive assistance in the development of this pipeline:

• @apeltzer
• @ewels
• @maxulysse
• @KevinMenden
• @bj-w

Acknowledgements

Contributions and Support

If you would like to contribute to this pipeline, please see the contributing guidelines.

For further information or help, don’t hesitate to get in touch on the Slack #circrna channel (you can join with this invite).

Citations

nf-core/circrna: a portable workflow for the quantification, miRNA target prediction and differential expression analysis of circular RNAs.

Barry Digby, Stephen P. Finn, & Pilib Ó Broin

BMC Bioinformatics 24, 27 (2023) doi: 10.1186/s12859-022-05125-8

An extensive list of references for the tools used by the pipeline can be found in the CITATIONS.md file.

You can cite the nf-core publication as follows:

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.