Package for mapping of GENCODE gene annotation files to older assemblies.

This provides tools to map the genome coordinates in GENCODE to previous releases of the reference genome. This projects annotations through genomics alignment using the TransMap algorithm to produce mappings. Reports are made on status of all mappings.

This maps all GENCODE GTF and GFF3 files. It does not map the PolyA_feature file. It is recommended to only use these on primary chromosomes, due to the high-similarity of alt-loci sequences to the primary assembly coupled with the growth in number of alt-loci making correct back-mapping difficult.

The program takes the current (source) GENCODE GFF3 or GTF, cross-assembly genomic alignments, and the previous (target) GENCODE annotations.

Mapping is done on a per-gene mapping using the following steps:

• Project transcripts of the gene through the alignments, keeping exons chained.
  • If there are multiple mappings, first look for ones that the overlapping to the previous version of the transcript, if it exists. Otherwise, if there is a previous version of the gene, select mappings overlapping the gene. Otherwise, to filter for paralog mappings, pick the mapping with the most similar span as the source.
  • Project features of the transcript, such as CDS and start codons, the transcript alignment between the genomes. This ensures that features stay in the same location within the transcript.
• Check all transcripts of the gene for consistency. Reject source gene mappings with transcripts on different chromosomes or strands, or where the genomic length of the gene has changed more than 50%.
• If a version of the gene exists in the target and the mapped gene doesn't overlap the target gene, it is also rejected.
  • If a gene did not map or was rejected and a version of the gene with the same biotype exists in the target annotations, use the existing gene.
• Small, automatic-only or all automatic genes are optionally not mapped, with the target annotation being passed through This avoids complex mappings of small RNAs imported from other database (e.g. mirRNAs).
• Target genes with no corresponding mappings and that overlap patched regions or regions with GRC indecent reports in the target genome may optionally be passed through. This address a fair number of problem cases. This was a common problem on GRCh37 chrX.

Pairing of source and target genes is somewhat complex due to instability of some gene identifiers between assemblies. If a matching base gene id (less version) is not found, an attempt is made to match the genes using the symbolic name.

Gene and transcript identifiers (*_id attribute) are based on the source identifiers. For annotations that are passed through unchanged from the target assembly, the identifiers are using as-is. For mapped annotations, the a mapping version number, in the form _n, is appended to the id. Where n is one based number indicating a version of the mapping. The mapping version start at 1' and are addition to the standard identifier version, for example ENST00000456328.2_1`

When a previous release file is supplied, the gene and transcript annotations are checked and the mapping version incremented if defined attributes change.

Genes have the mapping version numbers incremented if any of the following change from the previous release:

• contained transcripts, include version and mapping version number
• coordinates of the gene change
• the gene_type or gene_status changes
• the gene_name changes

Transcripts have the mapping version numbers incremented if any of the following change from the previous release:

• coordinates of the transcript or it's features change
• the transcript_type or transcript_status changes
• the transcript_name change
• any tag changes
• CDS or other sub-features of a transcript change.

The following ids have to the mapping versions applied

• gene_id
• transcript_id
• 'havana_gene
• 'havana_transcript

Information is collected on mappings and saved as attributes of the GFF3/GTF records. They are also records at the gene and transcript level the mapping information file. The attributes and their values are:

• remap_status - Attribute that indicates the status of the mapping. Possible values are:
  • full_contig - Gene or transcript completely mapped to the target genome with all features intact.
  • full_fragment - Gene or transcript completely to the target genome with insertions in some features. These are usually small insertions.
  • partial - Gene or transcript partially mapped to the target genome.
  • deleted - Gene or transcript did not map to the target genome.
  • no_seq_map - The source sequence is not in the assembly alignments. This will occur with alt loci genes if the alignments only contain the primary assembly.
  • gene_conflict - Transcripts in the gene mapped to multiple locations.
  • gene_size_change - Transcripts caused gene's length to change by more than 50%. This is to detect mapping to processed pseudogenes and mapping across tandem gene duplications.
  • automatic_small_ncrna_gene - Gene is a from a small, automatic (ENSEMBL source) non-coding RNA. These are take from the target annotations if --useTargetForAutoGenes is specified.
  • automatic_gene - Gene is a from an automatic process (ENSEMBL source). These are take from the target annotations if --useTargetForAutoGenes is specified.
  • pseudogene - Pseuduogene annotations (excluding polymorphic). These are take from the target annotations if --useTargetForPseudoGenes is specified.
• remap_original_id - Original ID attribute of the feature. If a feature is split when mapped, new IDs are created, otherwise the original ID is used.
• remap_original_location - Location of the feature in the source genome.
• remap_num_mappings - Number of mappings of the feature, only one of them was used.
• remap_target_status - Attribute that compares the mapping to the existing target annotations. Possible values are:
  • new - Gene or transcript was not in target annotations.
  • lost -Gene or transcript exists in source and target genome, however source was not mapped.
  • overlap - Gene or transcript overlaps previous version of annotation on target genome.
  • nonOverlap - Gene or transcript exists in target, however source mapping is to a different location. This is often mappings to a gene family members or pseudogenes.
• remap_substituted_missing_target - target gene annotate was substituted

The following files are needed to map using the UCSC liftover alignments:

• UCSC liftover alignments. These are the alignments of GRCh37 to GRCh38, which will be swapped. This produced slightly better results that using the
  • http://hgdownload.soe.ucsc.edu/goldenPath/hg38/liftOver/hg38ToHg19.over.chain.gz
• NCBI assembly report file for GRCh37
  • ftp://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/000/001/405/GCF_000001405.13_GRCh37/GCF_000001405.13_GRCh37_assembly_report.txt
• NCBI assembly report file for GRCh38. Since only the primary assembly is map, it is not necessary to update this for new releases.
  • ftp://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/000/001/405/GCF_000001405.36_GRCh38.p10/GCF_000001405.36_GRCh38.p10_assembly_report.txt
• GENCODE V19 primary annotations in GFF3 and GTF
  • ftp://ftp.sanger.ac.uk/pub/gencode/Gencode_human/release_19/gencode.v19.annotation.gtf.gz
  • ftp://ftp.sanger.ac.uk/pub/gencode/Gencode_human/release_19/gencode.v19.annotation.gff3.gz
• Current GENCODE primary assembly GFF3 and GTF files to map
  • ftp://ftp.sanger.ac.uk/pub/gencode/Gencode_human/release_25/gencode.v25.annotation.gff3.gz
  • ftp://ftp.sanger.ac.uk/pub/gencode/Gencode_human/release_25/gencode.v25.basic.annotation.gtf.gz
• GRC patch BED file for GRCh37 from UCSC. Download from
  • http://hgdownload.cse.ucsc.edu/goldenPath/hg19/database/altSeqPatchesP10.txt.gz
• GRC incident table from UCSC browser:
  • http://genomewiki.ucsc.edu/images/6/67/Hg19.grcIncidentDb.bb

Remove bin column from patches and convert BigBed to bed and combine

   zcat altSeqPatchesP10.txt.gz | cut -f 2- > altSeqPatchesP10.bed
   bigBedToBed Hg19.grcIncidentDb.bb Hg19.grcIncidentDb.bed
   cat altSeqPatchesP10.bed Hg19.grcIncidentDb.bed >problemRegions.bed

GENCODE uses different sequence naming for unplaced chromosome sequences than UCSC. Additionally, GENCODE annotates the GRCh37-lite chrM, while UCSC had incorporated a different chrM version. Convert the UCSC alignments to use GENCODE names:

../gencode-backmap/bin/ucscLiftEdit hg38ToHg19.over.chain.gz GCF_000001405.26_GRCh38_assembly_report.txt GCF_000001405.13_GRCh37_assembly_report.txt hg38ToHg19.over.gencode.chain

Map the annotation files:

../gencode-backmap/bin/gencode-backmap --swapMap --useTargetForAutoGenes --onlyManualForTargetSubstituteOverlap --substituteMissingTargets=V19 --headerFile=liftGxfHeader.txt --targetGxf=gencode.v19.annotation.gff3.gz --targetPatches=problemRegions.bed gencode.v25.annotation.gff3.gz  hg38ToHg19.over.gencode.chain gencode.v25lift37.annotation.gff3 gencode.v25lift37.map-info.tsv
../gencode-backmap/bin/gencode-backmap --swapMap --useTargetForAutoGenes --onlyManualForTargetSubstituteOverlap --substituteMissingTargets=V19 --headerFile=liftGxfHeader.txt --targetGxf=gencode.v19.annotation.gtf.gz --targetPatches=problemRegions.bed gencode.v25.annotation.gtf.gz  hg38ToHg19.over.gencode.chain gencode.v25lift37.annotation.gtf

Where liftGxfHeader.txt is the comments to add at the beginning of the output GFF3 or GTF files. This does not include GFF3 meta comment.

• GCC supporting -std=c++11 (4.7 or newer), or clang
• UCSC Browser library (kent/src)
• to run the tests, the following programs from the UCSC Browser are required:
  • gff3ToGenePred
  • gtfToGenePred
• Python 2.7

• Obtain and compile the libraries and command line utilities for the kent library:

  • http://genome-source.cse.ucsc.edu/gitweb/?p=kent.git;a=blob;f=src/product/README.building.source

• Edit config.mk to set the following variables

  • KENTDIR - point to kent/src directory
  • SAMTABIXDIR - if browser library is compiled with samtabix support, this is used to find the library.

• Compile code with make and turn tests with make test

• There is no install step, use directly from the bin directory

• The file src/version.h.in contains the version number. It should be update when doing a new release. The git sha1 hash of the commit is also added to this file. Be sure to do a make clean before doing a final build to get this file rebuild before a release.

• A tag is created in the form v1.0_rel on master. Due to the limited number of users of the code, release branches are not created.