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IsoformSwitchAnalyzeR Error in reading GTF file

Hi,

I am using IsoformSwitchAnalyzeR v1.21.0 in R 4.3.0 environment using RStudio (2023.06.0+421 "Mountain Hydrangea" Release). When I try to import GTF file (obtained from Ensembl Plants) I get the following error ;

Step 1 of 7: Checking data... Step 2 of 7: Obtaining annotation... importing GTF (this may take a while)... Error in dplyr::full_join(): ! ... must be empty.

I was using the following code;

### Create switchAnalyzeRlist
aSwitchList <- importRdata(
  isoformCountMatrix   = KallistoQuant$counts,
  isoformRepExpression = KallistoQuant$abundance,
  designMatrix         = myDesign,
  isoformExonAnnoation = "Arabidopsis_thaliana.TAIR10.53.gtf.gz",
  isoformNtFasta       = "Arabidopsis_thaliana.TAIR10.cdna.all.fa",
  comparisonsToMake = cond,
  showProgress = FALSE
)

Can anyone help me with this error? I have run this annotation file and fasta earlier without any issues (a year back).

Thank you in advance.

Venura

sessionInfo( )

R version 4.3.0 (2023-04-21 ucrt) Platform: x86_64-w64-mingw32/x64 (64-bit) Running under: Windows 10 x64 (build 19045)

Matrix products: default

locale: [1] LC_COLLATE=English_United States.utf8 LC_CTYPE=English_United States.utf8 LC_MONETARY=English_United States.utf8 LC_NUMERIC=C
[5] LC_TIME=English_United States.utf8

time zone: Asia/Colombo tzcode source: internal

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

other attached packages: [1] IsoformSwitchAnalyzeR_1.21.0 ggplot2_3.4.2 DEXSeq_1.46.0 RColorBrewer_1.1-3 AnnotationDbi_1.63.1
[6] DESeq2_1.41.2 SummarizedExperiment_1.31.1 GenomicRanges_1.53.1 GenomeInfoDb_1.37.2 IRanges_2.35.2
[11] S4Vectors_0.39.1 MatrixGenerics_1.13.0 matrixStats_1.0.0 Biobase_2.61.0 BiocGenerics_0.47.0
[16] BiocParallel_1.35.2 limma_3.57.6 BiocManager_1.30.21

loaded via a namespace (and not attached): [1] rstudioapi_0.14 jsonlite_1.8.5 tximport_1.29.0 magrittr_2.0.3 GenomicFeatures_1.53.1
[6] BiocIO_1.11.0 zlibbioc_1.47.0 vctrs_0.6.3 memoise_2.0.1 Rsamtools_2.17.0
[11] RCurl_1.98-1.12 htmltools_0.5.5 S4Arrays_1.1.4 progress_1.2.2 AnnotationHub_3.9.1
[16] lambda.r_1.2.4 curl_5.0.1 Rhdf5lib_1.23.0 rhdf5_2.45.0 SparseArray_1.1.10
[21] plyr_1.8.8 futile.options_1.0.1 cachem_1.0.8 GenomicAlignments_1.37.0 mime_0.12
[26] lifecycle_1.0.3 pkgconfig_2.0.3 Matrix_1.5-4.1 R6_2.5.1 fastmap_1.1.1
[31] GenomeInfoDbData_1.2.10 shiny_1.7.4 digest_0.6.31 colorspace_2.1-0 tximeta_1.19.0
[36] geneplotter_1.79.1 RSQLite_2.3.1 hwriter_1.3.2.1 filelock_1.0.2 fansi_1.0.4
[41] httr_1.4.6 compiler_4.3.0 bit64_4.0.5 withr_2.5.0 DBI_1.1.3
[46] biomaRt_2.57.1 rappdirs_0.3.3 DelayedArray_0.27.5 rjson_0.2.21 tools_4.3.0
[51] interactiveDisplayBase_1.39.0 DRIMSeq_1.29.0 httpuv_1.6.11 glue_1.6.2 VennDiagram_1.7.3
[56] restfulr_0.0.15 rhdf5filters_1.13.3 promises_1.2.0.1 grid_4.3.0 reshape2_1.4.4
[61] generics_0.1.3 gtable_0.3.3 BSgenome_1.69.0 tzdb_0.4.0 ensembldb_2.25.0
[66] hms_1.1.3 xml2_1.3.4 utf8_1.2.3 XVector_0.41.1 BiocVersion_3.18.0
[71] pillar_1.9.0 stringr_1.5.0 vroom_1.6.3 genefilter_1.83.1 later_1.3.1
[76] splines_4.3.0 dplyr_1.1.2 BiocFileCache_2.9.0 lattice_0.21-8 survival_3.5-5
[81] rtracklayer_1.61.0 bit_4.0.5 annotate_1.79.0 tidyselect_1.2.0 locfit_1.5-9.8
[86] Biostrings_2.69.1 gridExtra_2.3 ProtGenerics_1.33.1 edgeR_3.43.7 futile.logger_1.4.3
[91] statmod_1.5.0 stringi_1.7.12 lazyeval_0.2.2 yaml_2.3.7 codetools_0.2-19
[96] tibble_3.2.1 cli_3.6.1 xtable_1.8-4 munsell_0.5.0 Rcpp_1.0.10
[101] dbplyr_2.3.2 png_0.1-8 XML_3.99-0.14 parallel_4.3.0 ellipsis_0.3.2
[106] readr_2.1.4 blob_1.2.4 prettyunits_1.1.1 AnnotationFilter_1.25.0 bitops_1.0-7
[111] scales_1.2.1 crayon_1.5.2 rlang_1.1.1 KEGGREST_1.41.0 formatR_1.14
r isoformswitchanalyzer

Does this error persist when you gunzip your GTF file?

yes the same error appears.

My guess is that you are using cDNA sequence and not the unprocessed mRNA transcript.

1 answer

Hi All,

The error was version related. It was resolved once the developer version (instead of Bioconductor version) was installed using the following script.

if (!requireNamespace("devtools", quietly = TRUE)){
    install.packages("devtools")
}
devtools::install_github("kvittingseerup/IsoformSwitchAnalyzeR", build_vignettes = TRUE)

Thank you for all the assistance.

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