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I am trying to use IsoformSwitchAnalyzeR package to assess Isoform switching on mouse RNA seq data. I am running into problems on the GTF file that it needs to read in order to perform the analysis. I am not able to find the ensembl file that has haplotype info in it. Does anyone know where I can find this or how I would proceed using IsoformSwitchAnalyzeR without the "chr_patch_hapl_scaff.gtf" file.
Per my code below on the 'isoformExonAnnoation', I can only locate the 'Mus_musculus.GRCm39.107.chr.gtf' file, which it does not like and will not read in.
mySwitchList <- importRdata(
isoformCountMatrix = Txi_trans$counts,
isoformRepExpression = Txi_trans$abundance,
designMatrix = targets.mod,
removeNonConvensionalChr = TRUE,
addAnnotatedORFs=TRUE,
ignoreAfterPeriod=TRUE,
isoformExonAnnoation = "Mus_musculus.GRCm39.107.chr.gtf",
isoformNtFasta = "Mus_musculus.GRCm39.cdna.all.fa",
showProgress = TRUE)
sessionInfo()
R version 4.2.0 (2022-04-22)
Platform: x86_64-apple-darwin17.0 (64-bit)
Running under: macOS Big Sur 11.3
Matrix products: default
LAPACK: /Library/Frameworks/R.framework/Versions/4.2/Resources/lib/libRlapack.dylib
locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
attached base packages:
[1] stats4 stats graphics grDevices utils datasets methods base
other attached packages:
[1] IsoformSwitchAnalyzeR_1.18.0 DEXSeq_1.42.0 RColorBrewer_1.1-3
[4] DESeq2_1.36.0 SummarizedExperiment_1.26.1 MatrixGenerics_1.8.1
[7] BiocParallel_1.30.3 gt_0.6.0 plotly_4.10.0
[10] DT_0.23 cowplot_1.1.1 matrixStats_0.62.0
[13] edgeR_3.38.1 limma_3.52.2 beepr_1.3
[16] EnsDb.Mmusculus.v79_2.99.0 ensembldb_2.20.2 AnnotationFilter_1.20.0
[19] GenomicFeatures_1.48.3 AnnotationDbi_1.58.0 Biobase_2.56.0
[22] GenomicRanges_1.48.0 GenomeInfoDb_1.32.2 IRanges_2.30.0
[25] S4Vectors_0.34.0 BiocGenerics_0.42.0 tximport_1.24.0
[28] forcats_0.5.1 stringr_1.4.0 dplyr_1.0.9
[31] purrr_0.3.4 readr_2.1.2 tidyr_1.2.0
[34] tibble_3.1.8 ggplot2_3.3.6 tidyverse_1.3.2
[37] rhdf5_2.40.0 BiocManager_1.30.18
loaded via a namespace (and not attached):
[1] readxl_1.4.0 backports_1.4.1 AnnotationHub_3.4.0
[4] DRIMSeq_1.24.0 BiocFileCache_2.4.0 plyr_1.8.7
[7] lazyeval_0.2.2 tximeta_1.14.1 splines_4.2.0
[10] crosstalk_1.2.0 digest_0.6.29 htmltools_0.5.3
[13] fansi_1.0.3 magrittr_2.0.3 checkmate_2.1.0
[16] memoise_2.0.1 BSgenome_1.64.0 googlesheets4_1.0.0
[19] tzdb_0.3.0 annotate_1.74.0 Biostrings_2.64.0
[22] modelr_0.1.8 vroom_1.5.7 prettyunits_1.1.1
[25] colorspace_2.0-3 blob_1.2.3 rvest_1.0.2
[28] rappdirs_0.3.3 haven_2.5.0 crayon_1.5.1
[31] RCurl_1.98-1.7 jsonlite_1.8.0 hexbin_1.28.2
[34] genefilter_1.78.0 survival_3.3-1 glue_1.6.2
[37] gtable_0.3.0 gargle_1.2.0 zlibbioc_1.42.0
[40] XVector_0.36.0 DelayedArray_0.22.0 Rhdf5lib_1.18.2
[43] scales_1.2.0 futile.options_1.0.1 DBI_1.1.3
[46] Rcpp_1.0.9 xtable_1.8-4 viridisLite_0.4.0
[49] progress_1.2.2 bit_4.0.4 htmlwidgets_1.5.4
[52] httr_1.4.3 ellipsis_0.3.2 pkgconfig_2.0.3
[55] XML_3.99-0.10 farver_2.1.1 sass_0.4.2
[58] dbplyr_2.2.1 locfit_1.5-9.6 utf8_1.2.2
[61] later_1.3.0 reshape2_1.4.4 tidyselect_1.1.2
[64] labeling_0.4.2 rlang_1.0.4 BiocVersion_3.15.2
[67] munsell_0.5.0 cellranger_1.1.0 tools_4.2.0
[70] cachem_1.0.6 cli_3.3.0 generics_0.1.3
[73] RSQLite_2.2.15 audio_0.1-10 broom_1.0.0
[76] fastmap_1.1.0 yaml_2.3.5 bit64_4.0.5
[79] fs_1.5.2 KEGGREST_1.36.3 nlme_3.1-158
[82] mime_0.12 formatR_1.12 xml2_1.3.3
[85] biomaRt_2.52.0 compiler_4.2.0 rstudioapi_0.13
[88] interactiveDisplayBase_1.34.0 filelock_1.0.2 curl_4.3.2
[91] png_0.1-8 reprex_2.0.1 statmod_1.4.36
[94] geneplotter_1.74.0 bslib_0.4.0 stringi_1.7.8
[97] futile.logger_1.4.3 lattice_0.20-45 ProtGenerics_1.28.0
[100] Matrix_1.4-1 vctrs_0.4.1 pillar_1.8.0
[103] lifecycle_1.0.1 rhdf5filters_1.8.0 jquerylib_0.1.4
[106] data.table_1.14.2 bitops_1.0-7 httpuv_1.6.5
[109] rtracklayer_1.56.1 hwriter_1.3.2.1 R6_2.5.1
[112] BiocIO_1.6.0 promises_1.2.0.1 gridExtra_2.3
[115] sessioninfo_1.2.2 codetools_0.2-18 lambda.r_1.2.4
[118] MASS_7.3-58 assertthat_0.2.1 rjson_0.2.21
[121] withr_2.5.0 GenomicAlignments_1.32.1 Rsamtools_2.12.0
[124] GenomeInfoDbData_1.2.8 mgcv_1.8-40 parallel_4.2.0
[127] hms_1.1.1 VennDiagram_1.7.3 grid_4.2.0
[130] googledrive_2.0.0 shiny_1.7.2 lubridate_1.8.0
[133] restfulr_0.0.15
Hello Ryan, Also, I am not able to find esemble file that has haplotype info in it. Kindly, have you solved the problem?
Best
Which version of Ensembl are you using?