Problem with intronic variants in VariantAnnotation
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@paoloprovero-13328
Last seen 3.0 years ago
Italy

Hi I am using locateVariants from VariantAnnotation. Intronic variants seem to be associated to the wrong gene on a different chromosome

library(VariantAnnotation)
library(TxDb.Hsapiens.UCSC.hg19.knownGene)

variant <- GRanges(seqnames = "chr5", ranges = IRanges(start = 20298238
                                                       , end = 20298238))
genome(variant) <- "hg19"
anno <- locateVariants(query = variant, subject = TxDb.Hsapiens.UCSC.hg19.knownGene, region = AllVariants())
anno

GRanges object with 1 range and 9 metadata columns:
      seqnames    ranges strand | LOCATION  LOCSTART    LOCEND   QUERYID        TXID         CDSID      GENEID       PRECEDEID        FOLLOWID
         <Rle> <IRanges>  <Rle> | <factor> <integer> <integer> <integer> <character> <IntegerList> <character> <CharacterList> <CharacterList>
  [1]     chr5  20298238      - |   intron    277333    277333         1       19778                       839                                
  -------
  seqinfo: 1 sequence from an unspecified genome; no seqlengths

However the transcript identified by locateVariants is located on chromosome 4, not 5

dump <- as.list(TxDb.Hsapiens.UCSC.hg19.knownGene)
dump$transcripts[dump$transcripts$tx_id %in% anno$TXID,]   

      tx_id    tx_name tx_chrom tx_strand  tx_start    tx_end
19778 19778 uc003hzo.1     chr4         - 110609785 110624629

sessionInfo()

R version 4.1.2 (2021-11-01)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 20.04.3 LTS

Matrix products: default
BLAS:   /usr/lib/x86_64-linux-gnu/blas/libblas.so.3.9.0
LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.9.0

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C               LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8    LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C             LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] TxDb.Hsapiens.UCSC.hg19.knownGene_3.2.2 GenomicFeatures_1.44.0                 
 [3] AnnotationDbi_1.54.1                    VariantAnnotation_1.38.0               
 [5] Rsamtools_2.8.0                         Biostrings_2.60.1                      
 [7] XVector_0.32.0                          SummarizedExperiment_1.22.0            
 [9] Biobase_2.52.0                          GenomicRanges_1.44.0                   
[11] GenomeInfoDb_1.28.0                     IRanges_2.26.0                         
[13] S4Vectors_0.30.0                        MatrixGenerics_1.4.0                   
[15] matrixStats_0.61.0                      BiocGenerics_0.38.0                    

loaded via a namespace (and not attached):
 [1] Rcpp_1.0.7               lattice_0.20-45          prettyunits_1.1.1        png_0.1-7               
 [5] assertthat_0.2.1         digest_0.6.29            utf8_1.2.2               BiocFileCache_2.0.0     
 [9] R6_2.5.1                 RSQLite_2.2.9            httr_1.4.2               pillar_1.6.4            
[13] zlibbioc_1.38.0          rlang_0.4.12             progress_1.2.2           curl_4.3.2              
[17] rstudioapi_0.13          blob_1.2.2               Matrix_1.4-0             BiocParallel_1.26.0     
[21] stringr_1.4.0            RCurl_1.98-1.5           bit_4.0.4                biomaRt_2.48.1          
[25] DelayedArray_0.18.0      rtracklayer_1.52.0       compiler_4.1.2           pkgconfig_2.0.3         
[29] tidyselect_1.1.1         KEGGREST_1.32.0          tibble_3.1.6             GenomeInfoDbData_1.2.6  
[33] XML_3.99-0.8             fansi_0.5.0              crayon_1.4.2             dplyr_1.0.7             
[37] dbplyr_2.1.1             GenomicAlignments_1.28.0 bitops_1.0-7             rappdirs_0.3.3          
[41] grid_4.1.2               lifecycle_1.0.1          DBI_1.1.2                magrittr_2.0.1          
[45] stringi_1.7.6            cachem_1.0.6             xml2_1.3.3               ellipsis_0.3.2          
[49] filelock_1.0.2           vctrs_0.3.8              generics_0.1.1           rjson_0.2.20            
[53] restfulr_0.0.13          tools_4.1.2              bit64_4.0.5              BSgenome_1.60.0         
[57] glue_1.6.0               purrr_0.3.4              hms_1.1.1                yaml_2.2.1              
[61] fastmap_1.1.0            memoise_2.0.1            BiocIO_1.2.0

Thanks!

VariantAnnotation • 1.3k views
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There certainly seems to be a problem here. The TXID mapping seems problematic. Thanks for posting and we will get back to you.

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Is it clear what it would mean to associate an intronic variant with a TXID? The bug in locateVariants seems pretty clear to me -- a linear index is being treated as a string identifier, and we need to fix that. But I think the right answer for this location problem could be to return NA at TXID. The program does correctly say that the query GRanges is at an intron. [Edited to acknowledge my confusion. There could be multiple transcripts associated with an intronic variant and there is no reason not to list them all.]

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Hi, I have experienced exactly the same problem recently, are there any news about this issue? Many thanks!

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