Aaron Quinlan University of Utah quinlanlab.org @aaronquinlan ! ! ! ! Making queries of the genome less difficult. 

...CCTCATGCATGGAAA... Genetic variation ...CCTCATGTATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGTATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGTATGGAAA... Variant prioritization requires context. 

...CCTCATGCATGGAAA... Genetic variation ...CCTCATGTATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGTATGGAAA... ...CCTCATGCATGGAAA... ...CCTCATGTATGGAAA... Chromatin marks DNA methylation RNA expression TF binding Variant prioritization requires context. 

• inconsistent chromosome labels. • different sorting criteria. • mixed UNIX/Windows newlines. • file violates spec with vigor. • program expects exact extension. • file is gzipp’ed, not bgzipp’ed. • annotations use diff. genome builds. • tool only works for one format. • tool is hard-coded for specific build. • tool requires act of gods to compile. 

vcfanno will annotate your VCF with panache. Naked VCF vcfanno   +   configuration   file VCF w/ annotations in INFO field Brent Pedersen https://github.com/brentp/vcfanno VCF, BED, GFF, BAM, (soon BW) 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. Match on POS+REF+ALT for VCF annotations. 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. Allows multiple annotations from each file Match on POS+REF+ALT for VCF annotations. 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. Allows multiple annotations from each file Can rename the annotations in the resulting VCF Match on POS+REF+ALT for VCF annotations. 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. Allows multiple annotations from each file Can rename the annotations in the resulting VCF Multiple operations to summarize the results of multiple hits in annot. file: mean,  max,  min   concat,  count,  uniq   first,  flag Match on POS+REF+ALT for VCF annotations. 

[[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] vcfanno configuration file. Allows multiple annotations from each file Can rename the annotations in the resulting VCF Multiple operations to summarize the results of multiple hits in annot. file: mean,  max,  min   concat,  count,  uniq   first,  flag Match on POS+REF+ALT for VCF annotations. Javascript for custom computations. variance() defined in custom.js 

before and after vcfanno vcfanno AC=11;AF=0.017 AC=11;AF=0.017;   exac_aaf=0.0012;   exac_num_het=8;   rs_ids=1234;   gerp_mean=7.25e-­‐07   gerp_var=1.39e-­‐08 Naked VCF Dressed VCF [[annotation]]   file=“ExAC.v3.vcf”   fields=[“AF”,  “AC_Het”]   names=[“exac_aaf”,  “exac_num_het”]   ops=[“first”,  “first”]   ! [[annotation]]   file="dbsnp.b141.vcf.gz"   fields=["ID"]   names=["rs_ids"]   ops=[“concat"]   ! [[annotation]]   file="gerp.elements.bed.gz"   columns=[4,4]   names=[“gerp_mean”,”gerp_var”]   ops=[“mean”,  "js:variance(vals)"] 

New parallel “chromsweep”. vcfanno is speedy. 18 annotations: 29K variants / sec @ 12 cores See poster 160 for details 

Individual-centric queries with Genotype Query Tools (GQT) github.com/ryanlayer/gqt In press. Ryan Layer http://biorxiv.org/content/early/2015/04/20/018259 

A variant-centric query: Which variants affect BRCA1? 

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bcftools  view  \   -­‐r  17:43044295-­‐43125483  \   1000g.vcf                                      OR   ! tabix  1000g.vcf  17:43044295-­‐43125483   ! Existing tools handle variant-centric queries well 

An individual-centric query: In which variants are all affected males heterozygous? 

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In which variants are all affected males heterozygous? 

In which variants are all affected males heterozygous? 

Idea: transpose the genotype matrix G GT Note: other tricks included for speed/compression, please see manuscript 

In which variants are all affected males heterozygous? 

Affected males In which variants are all affected males heterozygous? 

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In which variants are all affected males heterozygous? 

In which variants are all affected males heterozygous? 

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Great, but what about indexing variant and genotype metadata? 

Bitmap indices of variant metadata (VEP consequence) VEP consequence bitmap: 1   0   0   0   0   0   0   …   0 synon. missense 0   0   0   0   0   0   1   …   0 stopgain 0   0   0   0   0   0   0   …   1 splice 0   0   0   0   1   0   0   …   0 .  .  . 

Bitmap indices of genotype metadata (depth) Ongoing: how to optimize lossiness of quantization? 

Bitmap indices of genotype metadata (depth) Genotype depth bitmap 1   0   0   0   0   0   0   …   0 0 1 0   1   0   0   0   0   0   …   0 2 0   0   0   0   0   0   1   …   0 3 0   0   0   1   0   0   0   …   0 10 0   0   0   0   0   0   0   …   1 20 0   0   0   0   0   1   0   …   0 25-30 0   0   0   0   1   0   0   …   0 >30 0   0   1   0   0   0   0   …   0 Ongoing: how to optimize lossiness of quantization? 

Future: A Genome Query Language? Variant-centric (bcftools, BGT) + = General Genome Query Language (based on discussions w/ Heng Li) VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in cases and rare in controls. b gqt query study.gqt study.db -p "phenotype == 2" -g "maf() > 0.05" -p "phenotype == 1" -g "maf() < 0.05" gqt -p -g b VCF In F V In VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in g Individual-centric (GQT, BGT) Individuals Variants Variants Individuals 

Future: A Genome Query Language? Variant-centric (bcftools, BGT) + = General Genome Query Language (based on discussions w/ Heng Li) VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in cases and rare in controls. b gqt query study.gqt study.db -p "phenotype == 2" -g "maf() > 0.05" -p "phenotype == 1" -g "maf() < 0.05" gqt -p -g b VCF In F V In VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in g Individual-centric (GQT, BGT) Individuals Variants Variants Individuals SELECT  *      VARIANT  gene="TP53"  AND  impact="HIGH"      SAMPLE  affected  IS  (ancestry="EA"                                                AND  phenotype=2                                              AND  BMI>35)      GENOTYPE  affected.MAF()>0.05 

Future: A Genome Query Language? Variant-centric (bcftools, BGT) + = General Genome Query Language (based on discussions w/ Heng Li) VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in cases and rare in controls. b gqt query study.gqt study.db -p "phenotype == 2" -g "maf() > 0.05" -p "phenotype == 1" -g "maf() < 0.05" gqt -p -g b VCF In F V In VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in g Individual-centric (GQT, BGT) Individuals Variants Variants Individuals SELECT  *      VARIANT  gene="TP53"  AND  impact="HIGH"      SAMPLE  affected  IS  (ancestry="EA"                                                AND  phenotype=2                                              AND  BMI>35)      GENOTYPE  affected.MAF()>0.05 

Future: A Genome Query Language? Variant-centric (bcftools, BGT) + = General Genome Query Language (based on discussions w/ Heng Li) VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in cases and rare in controls. b gqt query study.gqt study.db -p "phenotype == 2" -g "maf() > 0.05" -p "phenotype == 1" -g "maf() < 0.05" gqt -p -g b VCF In F V In VCF A B PED SQL database GQT index Individuals Variants 3 4 5 6 9 gqt convert ped gqt convert vcf D C Find variants that are common in g Individual-centric (GQT, BGT) Individuals Variants Variants Individuals SELECT  *      VARIANT  gene="TP53"  AND  impact="HIGH"      SAMPLE  affected  IS  (ancestry="EA"                                                AND  phenotype=2                                              AND  BMI>35)      GENOTYPE  affected.MAF()>0.05 

Thank you! Funding: Brent Pedersen Ryan Layer Jim Havrilla 

Students and Postdocs wanted. This could be you. Note: this is not me. [email protected]