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bio-sam-mutation

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bio-sam-mutationstveep/bioruby-sam-mutationHomepageDocumentationSource CodeBug TrackerWiki
Simple classes for parsing SAM, CIGAR and MD:Z strings, including slices. Methods for calling mutations in HGVS format and looking up consequences using Ensembl VEP REST API. Developed for calling mutations at an expected position in an alignment - e.g. Amplicon sequencing of CRISPR-induced mutations.
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 Popularity
Downloads
3,331
Stars
0
Forks
1
Watchers
3
 Releases
Current version
0.4.1
Total releases
1
First release
2016-02-08
Latest release
2016-02-08
 Issues
Open Issues
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Total Issues
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Issue Closure Rate
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 Development
Primary Language
Ruby
Licenses
MIT
Average date of last 50 commits
2016-05-15
Reverse Dependencies
0

 Dependencies

Development

bundler
>= 0
jeweler
~> 2.0
rdoc
~> 3.12
shoulda
>= 0
simplecov
>= 0
test-unit
~> 3.0

Runtime

bio
>= 1.4.2
bio-ensembl-rest
= 0.2.0
bio-samtools
~> 2.3.4
oj
~> 2.14
rake
~> 0.9
trollop
>= 0
 Project Readme

bio-sam-mutation

Build Status

  • Methods for calling mutations from SAM alignments, including CIGAR and MD tag parsers.

  • Annotates mutations in HGVS format: http://www.hgvs.org/mutnomen/recs.html.

  • Incorporates Ensembl VEP lookup.

Installation

gem install bio-sam-mutation

Usage

Command line

mutations -r -c config.yml *.sam

Produces a report using settings in config.yml. Report contains read counts for each mutant allele broken down by file. Use to identify mutants generated by CRISPR mutagenesis (for example), with settings to specify target region.

gem install bioruby-sam-mutation # Ruby > 2.1 required.

mutations -t -c config.yml *.sam

mutations --help

Tags a SAM file with mutation calls in HGVS format, using the settings in config.yml.

Example config file (YAML format). Multiple amplicons can be deconvoluted on-the-fly by specifiying the start sequence (allows reuse of sequencing barcodes). Transcript specifies the transcript of interest, required for Ensembl VEP consequence lookup. Translation start should be specified when aligning to cDNA sequence to get accurate HGVS annotation.

:species: "human"
:products:
  pool1:
    :start: "GGG"
    :offset: 25
    :length: 140
    :transcript: "ENST00000366794"
    :translation_start: 145
  pool2:
    :start: "AGT"
    :offset: 10
    :length: 320
    :transcript: "ENST00000366794"
    :translation_start: 145

Include colons prior to field names where shown (so they are interpreted as symbols in Ruby).

Gem

require 'bio-sam-mutation'

  # NB must be tab-delimited
  insertion_and_deletion = Bio::DB::Alignment.new("I2M5K:00253:00406\t0\t5\t112839854\t70\t63M2I138M1D27M7S\t*\t0\t0\tCAGTGATCTTCCAGATAGCCCTGGACAAACCATGCCACCAAGCAGAAGTAAAACACCTCCACCATACCTCCTCAAACAGCTCAAACCAAGCGAGAAGTACCTAAAAATAAAGCACCTACTGCTGAAAAGAGAGAGAGTGGACCTAAGCAAGCTGCAGTAAATGCTGCAGTTCAGAGGGTCCAGGTTCTTCCAGATGCTGATACTTATTACATTTTGCCACGGAAAGTACTGCTGAGG\t@CDDDCCCCACACCCCCCCC?CCACCCC>A6;;;;7;;6;6;BC;;6;;;;;.;;>ADDA??;;;;;?CCACCCD>C??@CCCC>C@C;>?CCCC@C=::@:::::+:::/:CCC?>>>>CCCCDDD9CCCC@AB????=AB>??;?BB>@@@AA???CC<@@?????BB>??;;;B<BC;??8;6:A=@=@BBB;;;?<77//*08*088888*8=9=?B7;;4;??????????<\tPG:Z:novoalign\tAS:i:183\tUQ:i:183\tNM:i:3\tMD:Z:201^T27")

  insertion_and_deletion.mutations
  #=> [#<Bio::Mutation:0x007fa20b5b4fc8 @position=112839916, @type=:insertion, @reference=nil, @mutant="AT", @seqname="5">, #<Bio::Mutation:0x007fa20b5b4960 @position=112840055, @type=:deletion, @reference="T", @mutant=nil, @seqname="5">]

  insertion_and_deletion.mutations.first.to_hgvs("g")
  #=> "5:g.112839916_112839917insAT"

  puts YAML.dump(insertion_and_deletion.mutations.first.vep("human","g").first["transcript_consequences"].keep_if{|c| c["transcript_id"] == "ENST00000257430"})
  #---
  # - variant_allele: AT
  #   cdna_end: 4379
  #   codons: cca/ccATa
  #   protein_end: 1441
  #   strand: 1
  #   hgnc_id: HGNC:583
  #   amino_acids: P/PX
  #   gene_symbol: APC
  #   cdna_start: 4378
  #   transcript_id: ENST00000257430
  #   cds_start: 4322
  #   gene_id: ENSG00000134982
  #   protein_start: 1441
  #   biotype: protein_coding
  #   gene_symbol_source: HGNC
  #   cds_end: 4323
  #   consequence_terms:
  #   - frameshift_variant
  #   impact: HIGH
  # => nil

# E.g. of full request return
# http://rest.ensembl.org/documentation/info/vep_hgvs_get
insertion_and_deletion.mutations(112839854).first.vep("human","g")
# => [{"assembly_name"=>"GRCh38", "end"=>112839917, "seq_region_name"=>"5", "transcript_consequences"=>[{"gene_id"=>"ENSG00000134982", "distance"=>46, "variant_allele"=>"AT", "biotype"=>"nonsense_mediated_decay", "gene_symbol_source"=>"HGNC", "consequence_terms"=>["downstream_gene_variant"], "strand"=>1, "hgnc_id"=>"HGNC:583", "gene_symbol"=>"APC", "transcript_id"=>"ENST00000502371", "impact"=>"MODIFIER"}, {"variant_allele"=>"AT", "cdna_end"=>4380, "codons"=>"-/AT", "protein_end"=>1442, "strand"=>1, "hgnc_id"=>"HGNC:583", "amino_acids"=>"-/X", "gene_symbol"=>"APC", "cdna_start"=>4379, "transcript_id"=>"ENST00000257430", "cds_start"=>4323, "gene_id"=>"ENSG00000134982", "protein_start"=>1441, "biotype"=>"protein_coding", "gene_symbol_source"=>"HGNC", "cds_end"=>4324, "consequence_terms"=>["frameshift_variant"], "impact"=>"HIGH"}, {"gene_id"=>"ENSG00000134982", "distance"=>863, "variant_allele"=>"AT", "biotype"=>"protein_coding", "gene_symbol_source"=>"HGNC", "consequence_terms"=>["downstream_gene_variant"], "strand"=>1, "hgnc_id"=>"HGNC:583", "gene_symbol"=>"APC", "transcript_id"=>"ENST00000507379", "impact"=>"MODIFIER"}, {"variant_allele"=>"AT", "cdna_end"=>4481, "codons"=>"-/AT", "protein_end"=>1442, "strand"=>1, "hgnc_id"=>"HGNC:583", "amino_acids"=>"-/X", "gene_symbol"=>"APC", "cdna_start"=>4480, "transcript_id"=>"ENST00000508376", "cds_start"=>4323, "gene_id"=>"ENSG00000134982", "protein_start"=>1441, "biotype"=>"protein_coding", "gene_symbol_source"=>"HGNC", "cds_end"=>4324, "consequence_terms"=>["frameshift_variant"], "impact"=>"HIGH"}, {"gene_id"=>"ENSG00000134982", "distance"=>409, "variant_allele"=>"AT", "biotype"=>"protein_coding", "gene_symbol_source"=>"HGNC", "consequence_terms"=>["downstream_gene_variant"], "strand"=>1, "hgnc_id"=>"HGNC:583", "gene_symbol"=>"APC", "transcript_id"=>"ENST00000512211", "impact"=>"MODIFIER"}, {"gene_id"=>"ENSG00000134982", "variant_allele"=>"AT", "cdna_end"=>4569, "biotype"=>"nonsense_mediated_decay", "gene_symbol_source"=>"HGNC", "consequence_terms"=>["3_prime_UTR_variant", "NMD_transcript_variant"], "strand"=>1, "hgnc_id"=>"HGNC:583", "gene_symbol"=>"APC", "cdna_start"=>4568, "transcript_id"=>"ENST00000508624", "impact"=>"MODIFIER"}, {"gene_id"=>"ENSG00000258864", "variant_allele"=>"AT", "biotype"=>"nonsense_mediated_decay", "gene_symbol_source"=>"Clone_based_vega_gene", "consequence_terms"=>["intron_variant", "NMD_transcript_variant"], "strand"=>1, "gene_symbol"=>"CTC-554D6.1", "transcript_id"=>"ENST00000520401", "impact"=>"MODIFIER"}, {"gene_id"=>"ENSG00000134982", "distance"=>2195, "variant_allele"=>"AT", "biotype"=>"protein_coding", "gene_symbol_source"=>"HGNC", "consequence_terms"=>["downstream_gene_variant"], "strand"=>1, "hgnc_id"=>"HGNC:583", "gene_symbol"=>"APC", "transcript_id"=>"ENST00000504915", "impact"=>"MODIFIER"}], "strand"=>1, "id"=>"5:g.112839917_112839918insAT", "allele_string"=>"-/AT", "most_severe_consequence"=>"frameshift_variant", "start"=>112839918}]

The API doc is online. For more code examples see the test files in the source tree.

Project home page

Information on the source tree, documentation, examples, issues and how to contribute, see

http://github.com/stveep/bioruby-sam-mutation

The BioRuby community is on IRC server: irc.freenode.org, channel: #bioruby.

Cite

If you use this software, please cite one of

Biogems.info

This Biogem is published at (http://biogems.info/index.html#bio-sam)

Copyright

Copyright (c) 2015 stveep. See LICENSE.txt for further details.