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Original file line number Diff line number Diff line change
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---
# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/environment-schema.json
channels:
- conda-forge
- bioconda
dependencies:
- "bioconda::leafcutter=2.0.3"
44 changes: 44 additions & 0 deletions modules/nf-core/leafcutter/differentialsplicing/main.nf
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process LEAFCUTTER_DIFFERENTIALSPLICING {
tag "$meta.id"
label 'process_medium'

conda "${moduleDir}/environment.yml"
container "${ workflow.containerEngine in ['singularity', 'apptainer'] && !task.ext.singularity_pull_docker_container ?
'https://depot.galaxyproject.org/singularity/leafcutter:2.0.3--pyhd8ed1ab_0':
'quay.io/biocontainers/leafcutter:2.0.3--pyhd8ed1ab_0' }"

input:
tuple val(meta), path(counts), path(groups)

output:
tuple val(meta), path("*_cluster_significance.txt"), emit: cluster_significance
tuple val(meta), path("*_effect_sizes.txt") , emit: effect_sizes
// WARN: Version information not provided by tool on CLI. Please update version string below when bumping container versions.
tuple val("${task.process}"), val('leafcutter'), val("2.0.3"), topic: versions, emit: versions_leafcutter

when:
task.ext.when == null || task.ext.when

script:
def args = task.ext.args ?: ''
def prefix = task.ext.prefix ?: "${meta.id}"
"""
export USER=\${USER:-nobody}
export PYTHONHASHSEED=0
export CUBLAS_WORKSPACE_CONFIG=:4096:8

leafcutter-ds \\
$counts \\
$groups \\
--output_prefix ${prefix}_results \\
--num_threads $task.cpus \\
$args
"""

stub:
def prefix = task.ext.prefix ?: "${meta.id}"
"""
touch ${prefix}_results_cluster_significance.txt
touch ${prefix}_results_effect_sizes.txt
"""
}
80 changes: 80 additions & 0 deletions modules/nf-core/leafcutter/differentialsplicing/meta.yml
Original file line number Diff line number Diff line change
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# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/meta-schema.json
name: "leafcutter_differentialsplicing"
description: Perform differential splicing analysis to identify junctions with significant changes in splicing between conditions
keywords:
- alternative splicing
- differential splicing
- RNA-seq
- intron usage
tools:
- "leafcutter":
description: "Annotation-free quantification of RNA splicing from RNA-seq data"
homepage: "https://leafcutter2.github.io/leafcutter-ds/"
documentation: "https://leafcutter2.github.io/leafcutter-ds/tutorials/differential-splicing/"
tool_dev_url: "https://github.com/leafcutter2/leafcutter-ds"
doi: "10.1038/s41588-017-0004-9"
licence: ["Apache-2.0"]
identifier: biotools:leafcutter

input:
- - meta:
type: map
description: |
Groovy Map containing sample information
e.g. `[ id:'test' ]`
- counts:
type: file
description: Intron usage counts file from leafcutter-cluster (either perind.counts.gz or perind_numers.counts.gz)
pattern: "*.counts.gz"
- groups:
type: file
description: Tab-delimited file with sample names in column 1 and sample group/phenotype assignments in column 2
pattern: "*.txt"

output:
cluster_significance:
- - meta:
type: map
description: |
Groovy Map containing sample information
e.g. `[ id:'test' ]`
- "*_cluster_significance.txt":
type: file
description: Cluster-level differential splicing statistics including p-values and test status
pattern: "*_cluster_significance.txt"
effect_sizes:
- - meta:
type: map
description: |
Groovy Map containing sample information
e.g. `[ id:'test' ]`
- "*_effect_sizes.txt":
type: file
description: Per-intron junction effect sizes, PSI values, and delta PSI for each condition
pattern: "*_effect_sizes.txt"
versions_leafcutter:
- - "${task.process}":
type: string
description: The name of the process
- "leafcutter":
type: string
description: The name of the tool
- "2.0.3":
type: string
description: The version of the tool

topics:
versions:
- - ${task.process}:
type: string
description: The name of the process
- leafcutter:
type: string
description: The name of the tool
- "2.0.3":
type: string
description: The version of the tool
authors:
- "@JTL-lab"
maintainers:
- "@JTL-lab"
124 changes: 124 additions & 0 deletions modules/nf-core/leafcutter/differentialsplicing/tests/main.nf.test
Original file line number Diff line number Diff line change
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nextflow_process {

name "Test Process LEAFCUTTER_DIFFERENTIALSPLICING"
script "../main.nf"
process "LEAFCUTTER_DIFFERENTIALSPLICING"

tag "modules"
tag "modules_nfcore"
tag "leafcutter"
tag "leafcutter/differentialsplicing"
tag "leafcutter/clusterregtools"
tag "regtools"
tag "regtools/junctionsextract"

test("homo_sapiens - bam - differential splicing") {

config "./nextflow.config"

setup {
run("REGTOOLS_JUNCTIONSEXTRACT") {
script "../../../regtools/junctionsextract/main.nf"
process {
"""
input[0] = Channel.of(
[
[ id:'ctrl_rep1', single_end:false ],
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam", checkIfExists: true),
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam.bai", checkIfExists: true)
],
[
[ id:'ctrl_rep2', single_end:false ],
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam", checkIfExists: true),
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam.bai", checkIfExists: true)
],
[
[ id:'kd_rep1', single_end:false ],
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam", checkIfExists: true),
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam.bai", checkIfExists: true)
],
[
[ id:'kd_rep2', single_end:false ],
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam", checkIfExists: true),
file(params.modules_testdata_base_path + "genomics/homo_sapiens/illumina/bam/test.rna.paired_end.sorted.chr6.bam.bai", checkIfExists: true)
]
)
input[1] = ''
"""
}
}

run("LEAFCUTTER_CLUSTERREGTOOLS") {
script "../../../leafcutter/clusterregtools/main.nf"
process {
"""
input[0] = REGTOOLS_JUNCTIONSEXTRACT.out.junc
.map { meta, junc -> junc }
.collect()
.map { junc_files -> [ [ id:'test_ds' ], junc_files ] }
"""
}
}
}

when {
process {
"""
def groups_content = '''ctrl_rep1\\tCONTROL
ctrl_rep2\\tCONTROL
kd_rep1\\tKNOCKDOWN
kd_rep2\\tKNOCKDOWN
'''
def groups_file = file("\${workDir}/groups_file.txt")
groups_file.text = groups_content

input[0] = LEAFCUTTER_CLUSTERREGTOOLS.out.numers
.map { meta, counts -> [ meta, counts, groups_file ] }
"""
}
}

then {
assertAll(
{ assert process.success },
{ assert snapshot(
process.out.findAll { key, val -> key.startsWith('versions') },
process.out.cluster_significance.collect { file(it[1]).getName() },
process.out.effect_sizes.collect { file(it[1]).getName() }
).match() },
// Check non-deterministic files exist
{ assert file(process.out.cluster_significance[0][1]).exists() },
{ assert file(process.out.effect_sizes[0][1]).exists() }
)
}

}

test("homo_sapiens - bam - differential splicing - stub") {

options "-stub"

when {
process {
"""
input[0] = [
[ id:'test' ],
file('test_perind.counts.gz'),
file('groups_file.txt')
]
"""
}
}

then {
assertAll(
{ assert process.success },
{ assert snapshot(process.out).match() },
{ assert process.out.cluster_significance != null },
{ assert process.out.effect_sizes != null }
)
}

}

}
Original file line number Diff line number Diff line change
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{
"homo_sapiens - bam - differential splicing": {
"content": [
{
"versions_leafcutter": [
[
"LEAFCUTTER_DIFFERENTIALSPLICING",
"leafcutter",
"2.0.3"
]
]
},
[
"test_ds_results_cluster_significance.txt"
],
[
"test_ds_results_effect_sizes.txt"
]
],
"timestamp": "2026-07-08T14:38:48.580335473",
"meta": {
"nf-test": "0.9.5",
"nextflow": "25.10.0"
}
},
"homo_sapiens - bam - differential splicing - stub": {
"content": [
{
"0": [
[
{
"id": "test"
},
"test_results_cluster_significance.txt:md5,d41d8cd98f00b204e9800998ecf8427e"
]
],
"1": [
[
{
"id": "test"
},
"test_results_effect_sizes.txt:md5,d41d8cd98f00b204e9800998ecf8427e"
]
],
"2": [
[
"LEAFCUTTER_DIFFERENTIALSPLICING",
"leafcutter",
"2.0.3"
]
],
"cluster_significance": [
[
{
"id": "test"
},
"test_results_cluster_significance.txt:md5,d41d8cd98f00b204e9800998ecf8427e"
]
],
"effect_sizes": [
[
{
"id": "test"
},
"test_results_effect_sizes.txt:md5,d41d8cd98f00b204e9800998ecf8427e"
]
],
"versions_leafcutter": [
[
"LEAFCUTTER_DIFFERENTIALSPLICING",
"leafcutter",
"2.0.3"
]
]
}
],
"timestamp": "2026-07-08T14:39:52.087856717",
"meta": {
"nf-test": "0.9.5",
"nextflow": "25.10.0"
}
}
}
Original file line number Diff line number Diff line change
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process {

withName: 'LEAFCUTTER_DIFFERENTIALSPLICING' {
ext.args = '--min_samples_per_group 2 --min_samples_per_intron 2 --num_threads 1'
}

}