From 901fabf8aad66fe8f3563dd3d3d65e6acaeb3a7a Mon Sep 17 00:00:00 2001 From: Lucille Delisle Date: Wed, 19 Jun 2024 07:33:53 +0200 Subject: [PATCH 1/2] Add brew3r --- .../transcriptomics/brew3r/.dockstore.yml | 11 + .../transcriptomics/brew3r/BREW3R-tests.yml | 29 ++ workflows/transcriptomics/brew3r/BREW3R.ga | 455 ++++++++++++++++++ workflows/transcriptomics/brew3r/CHANGELOG.md | 5 + workflows/transcriptomics/brew3r/README.md | 26 + .../brew3r/test-data/GSM4196550_subset.bam | Bin 0 -> 100331 bytes .../brew3r/test-data/GSM4196551_subset.bam | Bin 0 -> 151588 bytes .../brew3r/test-data/GSM4196552_subset.bam | Bin 0 -> 169186 bytes .../brew3r/test-data/input.gtf | 8 + 9 files changed, 534 insertions(+) create mode 100644 workflows/transcriptomics/brew3r/.dockstore.yml create mode 100644 workflows/transcriptomics/brew3r/BREW3R-tests.yml create mode 100644 workflows/transcriptomics/brew3r/BREW3R.ga create mode 100644 workflows/transcriptomics/brew3r/CHANGELOG.md create mode 100644 workflows/transcriptomics/brew3r/README.md create mode 100644 workflows/transcriptomics/brew3r/test-data/GSM4196550_subset.bam create mode 100644 workflows/transcriptomics/brew3r/test-data/GSM4196551_subset.bam create mode 100644 workflows/transcriptomics/brew3r/test-data/GSM4196552_subset.bam create mode 100644 workflows/transcriptomics/brew3r/test-data/input.gtf diff --git a/workflows/transcriptomics/brew3r/.dockstore.yml b/workflows/transcriptomics/brew3r/.dockstore.yml new file mode 100644 index 000000000..37383bd43 --- /dev/null +++ b/workflows/transcriptomics/brew3r/.dockstore.yml @@ -0,0 +1,11 @@ +version: 1.2 +workflows: +- name: main + subclass: Galaxy + publish: true + primaryDescriptorPath: /BREW3R.ga + testParameterFiles: + - /BREW3R-tests.yml + authors: + - name: Lucille Delisle + orcid: 0000-0002-1964-4960 diff --git a/workflows/transcriptomics/brew3r/BREW3R-tests.yml b/workflows/transcriptomics/brew3r/BREW3R-tests.yml new file mode 100644 index 000000000..77be3b2df --- /dev/null +++ b/workflows/transcriptomics/brew3r/BREW3R-tests.yml @@ -0,0 +1,29 @@ +- doc: Test outline for BREW3R + job: + Input gtf: + class: File + path: test-data/input.gtf + filetype: gtf + BAM collection: + class: Collection + collection_type: list + elements: + - class: File + identifier: GSM4196550 + path: test-data/GSM4196550_subset.bam + - class: File + identifier: GSM4196551 + path: test-data/GSM4196551_subset.bam + - class: File + identifier: GSM4196552 + path: test-data/GSM4196552_subset.bam + strandedness: 'stranded - reverse' + minimum coverage: '10' + minimum FPKM for merge: '1.0' + outputs: + extended_gtf: + asserts: + has_line: + line: 'chr12 HAVANA exon 11206402 11208972 . - . gene_id "ENSMUSG00000047002.2"; gene_type "protein_coding"; gene_name "Msgn1"; level "2"; mgi_id "MGI:1860483"; havana_gene "OTTMUSG00000065713.1"; transcript_id "ENSMUST00000049877.2"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_transcript "OTTMUST00000159929.1"; exon_number "1"; exon_id "ENSMUSE00000372682.2.ext"' + has_text: + text: "CDS" diff --git a/workflows/transcriptomics/brew3r/BREW3R.ga b/workflows/transcriptomics/brew3r/BREW3R.ga new file mode 100644 index 000000000..1c035db24 --- /dev/null +++ b/workflows/transcriptomics/brew3r/BREW3R.ga @@ -0,0 +1,455 @@ +{ + "a_galaxy_workflow": "true", + "annotation": "This workflow takes a collection of BAM (output of STAR) and a gtf. 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a/workflows/transcriptomics/brew3r/README.md b/workflows/transcriptomics/brew3r/README.md new file mode 100644 index 000000000..1430f2262 --- /dev/null +++ b/workflows/transcriptomics/brew3r/README.md @@ -0,0 +1,26 @@ +# BREW3R + +![BREW3R logo](https://raw.githubusercontent.com/lldelisle/BREW3R/main/images/logo.png) + +BREW3R stands for **B**ulk **R**NA-seq **E**vidence-based **W**orkflow for **3**' UTR **R**eannotation. + +This workflow enables extending an existing gtf downloaded on a public website, like Ensembl, Genecode or UCSC, using *de novo* gene annotation with StringTie on full length bulk RNA-seq. + +BREW3R highly relies on a R package called BREW3R.r available on [bioconductor](https://bioconductor.org/packages/release/bioc/html/BREW3R.r.html). + +## Input datasets + +- The workflow requires an input gtf file which will be extended. +- As well as a collection of BAM files. + +## Input values + +- strandedness: Must be one of `stranded - forward`, `stranded - reverse` and `unstranded`. For stranded libraries, reverse means that the read is complementary to the coding sequence, forward means that the read is in the same orientation as the coding sequence. +- minimum coverage: Minimum reads per bp coverage to consider for assembly in each de novo assembly (for each BAM file). Default: 10 +- minimum FPKM for merge: Minimum FPKM value for a transcript to be included into the merged de novo assembly. + +## Processing + +- StringTie is called once per input BAM file to compute de novo assembly. +- StringTie is called to merge all outputs of previous steps. +- BREW3R.r is run with the default parameters on the input gtf to extend and the output of StringTie. 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a/workflows/transcriptomics/brew3r/test-data/input.gtf b/workflows/transcriptomics/brew3r/test-data/input.gtf new file mode 100644 index 000000000..ef0e00175 --- /dev/null +++ b/workflows/transcriptomics/brew3r/test-data/input.gtf @@ -0,0 +1,8 @@ +chr12 HAVANA gene 11208360 11208972 . - . gene_id "ENSMUSG00000047002.2"; gene_type "protein_coding"; gene_name "Msgn1"; level 2; mgi_id "MGI:1860483"; havana_gene "OTTMUSG00000065713.1"; +chr12 HAVANA transcript 11208360 11208972 . - . gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA exon 11208360 11208972 . - . gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA CDS 11208385 11208948 . - 0 gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA start_codon 11208946 11208948 . - 0 gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA stop_codon 11208382 11208384 . - 0 gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA UTR 11208360 11208384 . - . gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; +chr12 HAVANA UTR 11208949 11208972 . - . gene_id "ENSMUSG00000047002.2"; transcript_id "ENSMUST00000049877.2"; gene_type "protein_coding"; gene_name "Msgn1"; transcript_type "protein_coding"; transcript_name "Msgn1-201"; exon_number 1; exon_id "ENSMUSE00000372682.2"; level 2; protein_id "ENSMUSP00000055001.1"; transcript_support_level "NA"; mgi_id "MGI:1860483"; tag "basic"; tag "appris_principal_1"; tag "CCDS"; ccdsid "CCDS25814.1"; havana_gene "OTTMUSG00000065713.1"; havana_transcript "OTTMUST00000159929.1"; From 44561d7bd9ab8d91a09c97a369c36d1553a53530 Mon Sep 17 00:00:00 2001 From: Lucille Delisle Date: Wed, 19 Jun 2024 09:15:59 +0200 Subject: [PATCH 2/2] Remove quotes around int/float parameter values Co-authored-by: Marius van den Beek --- workflows/transcriptomics/brew3r/BREW3R-tests.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/transcriptomics/brew3r/BREW3R-tests.yml b/workflows/transcriptomics/brew3r/BREW3R-tests.yml index 77be3b2df..9e699b3d8 100644 --- a/workflows/transcriptomics/brew3r/BREW3R-tests.yml +++ b/workflows/transcriptomics/brew3r/BREW3R-tests.yml @@ -18,8 +18,8 @@ identifier: GSM4196552 path: test-data/GSM4196552_subset.bam strandedness: 'stranded - reverse' - minimum coverage: '10' - minimum FPKM for merge: '1.0' + minimum coverage: 10 + minimum FPKM for merge: 1.0 outputs: extended_gtf: asserts: