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Visualization

  1. Load the data to the UCSC (IGV) browser to visualize it.
#Use the bed files from the tracks and tables folder
# Check for the gene LIG4
  1. Add additional tracks, like functional regions from ENCODE, TFBSs, etc

Location

  1. Annotate regions using the annotatr package
#file <- fread(file="data/diffMethTable_site_cmp1.csv", sep = ",")


  #bg <- copy(file)
  #bg <- bg[,.(Chromosome, Start, End=Start+1, cgid)]
  #fwrite(bg, row.names=F, file = "rnb_run_pilot/differential_methylation_data/background.txt", quote = F, sep="\t",  col.names = F)

#hyper <- file[diffmeth.p.val<0.0001 & mean.diff>0.3,]
#great_hyper <- hyper[,.(Chromosome, Start, End=Start+1, cgid)]

#hypo <- file[diffmeth.p.val<10e-14 & mean.diff< -0.7,]
#great_hypo <- hypo[,.(Chromosome, Start, End=Start+1, cgid)]

#fwrite(file, row.names=F, file = paste0(folder, "diffMethTable_significant01_cmp_", i, ".txt"), quote = F, sep="\t")

#fwrite(great_hyper, row.names=F, file = "rnb_run_pilot/differential_methylation_data/great_significant_hyper.txt", quote = F, sep="\t", col.names = F)
#fwrite(great_hypo, row.names=F, file = "rnb_run_pilot/differential_methylation_data/great_significant_hypo.txt", quote = F, sep="\t", col.names = F)



data <- list()
data[["hypo"]]  <- import.bed("../inst/extdata/great_significant_hypo.txt")
data[["hyper"]]<- import.bed("../inst/extdata/great_significant_hyper.txt")


annots = c('hg19_Hepg2-chromatin')
annots_gr = build_annotations(genome = 'hg19', annotations = annots)
## Building hmms...
## Downloading chromHMM track for Hepg2
for (dataset in data){
genome(dataset) <- rep("hg19", length(genome(dataset)))
result <- annotate_regions(dataset, annotations=annots_gr, minoverlap = 1L, ignore.strand = TRUE, quiet = FALSE)
rnd_annots = annotate_regions(regions = randomize_regions(dataset),annotations = annots_gr,ignore.strand = TRUE)
p_annots = plot_annotation(annotated_regions = result, annotated_random = rnd_annots)
print(p_annots)
}
## Annotating...
## Randomizing regions...
## Annotating...
## Annotating...
## Randomizing regions...
## Annotating...

background <-  import.bed("../inst/extdata/background.txt")
genome(background) <- rep("hg19", length(genome(background)))


for (dataset in names(data)){
genome(data[[dataset]]) <- rep("hg19", length(genome(data[[dataset]])))
background2 <- background[sample(1:length(background), length(data[[dataset]])),]
result <- annotate_regions(data[[dataset]], annotations=annots_gr, minoverlap = 1L, ignore.strand = TRUE, quiet = FALSE)
rnd_annots = annotate_regions(regions = background2,annotations = annots_gr,ignore.strand = TRUE)
p_annots = plot_annotation(annotated_regions = result, annotated_random = rnd_annots)
print(p_annots+ggtitle(dataset))
}
## Annotating...
## Annotating...
## Annotating...
## Annotating...

  1. Try different annotations
builtin_annotations()
##   [1] "dm3_genes_1to5kb"                     
##   [2] "dm6_genes_1to5kb"                     
##   [3] "galGal5_genes_1to5kb"                 
##   [4] "hg19_genes_1to5kb"                    
##   [5] "hg38_genes_1to5kb"                    
##   [6] "mm9_genes_1to5kb"                     
##   [7] "mm10_genes_1to5kb"                    
##   [8] "rn4_genes_1to5kb"                     
##   [9] "rn5_genes_1to5kb"                     
##  [10] "rn6_genes_1to5kb"                     
##  [11] "dm3_genes_promoters"                  
##  [12] "dm6_genes_promoters"                  
##  [13] "galGal5_genes_promoters"              
##  [14] "hg19_genes_promoters"                 
##  [15] "hg38_genes_promoters"                 
##  [16] "mm9_genes_promoters"                  
##  [17] "mm10_genes_promoters"                 
##  [18] "rn4_genes_promoters"                  
##  [19] "rn5_genes_promoters"                  
##  [20] "rn6_genes_promoters"                  
##  [21] "dm3_genes_cds"                        
##  [22] "dm6_genes_cds"                        
##  [23] "galGal5_genes_cds"                    
##  [24] "hg19_genes_cds"                       
##  [25] "hg38_genes_cds"                       
##  [26] "mm9_genes_cds"                        
##  [27] "mm10_genes_cds"                       
##  [28] "rn4_genes_cds"                        
##  [29] "rn5_genes_cds"                        
##  [30] "rn6_genes_cds"                        
##  [31] "dm3_genes_5UTRs"                      
##  [32] "dm6_genes_5UTRs"                      
##  [33] "galGal5_genes_5UTRs"                  
##  [34] "hg19_genes_5UTRs"                     
##  [35] "hg38_genes_5UTRs"                     
##  [36] "mm9_genes_5UTRs"                      
##  [37] "mm10_genes_5UTRs"                     
##  [38] "rn4_genes_5UTRs"                      
##  [39] "rn5_genes_5UTRs"                      
##  [40] "rn6_genes_5UTRs"                      
##  [41] "dm3_genes_exons"                      
##  [42] "dm6_genes_exons"                      
##  [43] "galGal5_genes_exons"                  
##  [44] "hg19_genes_exons"                     
##  [45] "hg38_genes_exons"                     
##  [46] "mm9_genes_exons"                      
##  [47] "mm10_genes_exons"                     
##  [48] "rn4_genes_exons"                      
##  [49] "rn5_genes_exons"                      
##  [50] "rn6_genes_exons"                      
##  [51] "dm3_genes_firstexons"                 
##  [52] "dm6_genes_firstexons"                 
##  [53] "galGal5_genes_firstexons"             
##  [54] "hg19_genes_firstexons"                
##  [55] "hg38_genes_firstexons"                
##  [56] "mm9_genes_firstexons"                 
##  [57] "mm10_genes_firstexons"                
##  [58] "rn4_genes_firstexons"                 
##  [59] "rn5_genes_firstexons"                 
##  [60] "rn6_genes_firstexons"                 
##  [61] "dm3_genes_introns"                    
##  [62] "dm6_genes_introns"                    
##  [63] "galGal5_genes_introns"                
##  [64] "hg19_genes_introns"                   
##  [65] "hg38_genes_introns"                   
##  [66] "mm9_genes_introns"                    
##  [67] "mm10_genes_introns"                   
##  [68] "rn4_genes_introns"                    
##  [69] "rn5_genes_introns"                    
##  [70] "rn6_genes_introns"                    
##  [71] "dm3_genes_intronexonboundaries"       
##  [72] "dm6_genes_intronexonboundaries"       
##  [73] "galGal5_genes_intronexonboundaries"   
##  [74] "hg19_genes_intronexonboundaries"      
##  [75] "hg38_genes_intronexonboundaries"      
##  [76] "mm9_genes_intronexonboundaries"       
##  [77] "mm10_genes_intronexonboundaries"      
##  [78] "rn4_genes_intronexonboundaries"       
##  [79] "rn5_genes_intronexonboundaries"       
##  [80] "rn6_genes_intronexonboundaries"       
##  [81] "dm3_genes_exonintronboundaries"       
##  [82] "dm6_genes_exonintronboundaries"       
##  [83] "galGal5_genes_exonintronboundaries"   
##  [84] "hg19_genes_exonintronboundaries"      
##  [85] "hg38_genes_exonintronboundaries"      
##  [86] "mm9_genes_exonintronboundaries"       
##  [87] "mm10_genes_exonintronboundaries"      
##  [88] "rn4_genes_exonintronboundaries"       
##  [89] "rn5_genes_exonintronboundaries"       
##  [90] "rn6_genes_exonintronboundaries"       
##  [91] "dm3_genes_3UTRs"                      
##  [92] "dm6_genes_3UTRs"                      
##  [93] "galGal5_genes_3UTRs"                  
##  [94] "hg19_genes_3UTRs"                     
##  [95] "hg38_genes_3UTRs"                     
##  [96] "mm9_genes_3UTRs"                      
##  [97] "mm10_genes_3UTRs"                     
##  [98] "rn4_genes_3UTRs"                      
##  [99] "rn5_genes_3UTRs"                      
## [100] "rn6_genes_3UTRs"                      
## [101] "dm3_genes_intergenic"                 
## [102] "dm6_genes_intergenic"                 
## [103] "galGal5_genes_intergenic"             
## [104] "hg19_genes_intergenic"                
## [105] "hg38_genes_intergenic"                
## [106] "mm9_genes_intergenic"                 
## [107] "mm10_genes_intergenic"                
## [108] "rn4_genes_intergenic"                 
## [109] "rn5_genes_intergenic"                 
## [110] "rn6_genes_intergenic"                 
## [111] "galGal5_cpg_islands"                  
## [112] "hg19_cpg_islands"                     
## [113] "hg38_cpg_islands"                     
## [114] "mm9_cpg_islands"                      
## [115] "mm10_cpg_islands"                     
## [116] "rn4_cpg_islands"                      
## [117] "rn5_cpg_islands"                      
## [118] "rn6_cpg_islands"                      
## [119] "galGal5_cpg_shores"                   
## [120] "hg19_cpg_shores"                      
## [121] "hg38_cpg_shores"                      
## [122] "mm9_cpg_shores"                       
## [123] "mm10_cpg_shores"                      
## [124] "rn4_cpg_shores"                       
## [125] "rn5_cpg_shores"                       
## [126] "rn6_cpg_shores"                       
## [127] "galGal5_cpg_shelves"                  
## [128] "hg19_cpg_shelves"                     
## [129] "hg38_cpg_shelves"                     
## [130] "mm9_cpg_shelves"                      
## [131] "mm10_cpg_shelves"                     
## [132] "rn4_cpg_shelves"                      
## [133] "rn5_cpg_shelves"                      
## [134] "rn6_cpg_shelves"                      
## [135] "galGal5_cpg_inter"                    
## [136] "hg19_cpg_inter"                       
## [137] "hg38_cpg_inter"                       
## [138] "mm9_cpg_inter"                        
## [139] "mm10_cpg_inter"                       
## [140] "rn4_cpg_inter"                        
## [141] "rn5_cpg_inter"                        
## [142] "rn6_cpg_inter"                        
## [143] "hg19_chromatin_Gm12878-ActivePromoter"
## [144] "hg19_chromatin_H1hesc-ActivePromoter" 
## [145] "hg19_chromatin_Hepg2-ActivePromoter"  
## [146] "hg19_chromatin_Hmec-ActivePromoter"   
## [147] "hg19_chromatin_Hsmm-ActivePromoter"   
## [148] "hg19_chromatin_Huvec-ActivePromoter"  
## [149] "hg19_chromatin_K562-ActivePromoter"   
## [150] "hg19_chromatin_Nhek-ActivePromoter"   
## [151] "hg19_chromatin_Nhlf-ActivePromoter"   
## [152] "hg19_chromatin_Gm12878-WeakPromoter"  
## [153] "hg19_chromatin_H1hesc-WeakPromoter"   
## [154] "hg19_chromatin_Hepg2-WeakPromoter"    
## [155] "hg19_chromatin_Hmec-WeakPromoter"     
## [156] "hg19_chromatin_Hsmm-WeakPromoter"     
## [157] "hg19_chromatin_Huvec-WeakPromoter"    
## [158] "hg19_chromatin_K562-WeakPromoter"     
## [159] "hg19_chromatin_Nhek-WeakPromoter"     
## [160] "hg19_chromatin_Nhlf-WeakPromoter"     
## [161] "hg19_chromatin_Gm12878-PoisedPromoter"
## [162] "hg19_chromatin_H1hesc-PoisedPromoter" 
## [163] "hg19_chromatin_Hepg2-PoisedPromoter"  
## [164] "hg19_chromatin_Hmec-PoisedPromoter"   
## [165] "hg19_chromatin_Hsmm-PoisedPromoter"   
## [166] "hg19_chromatin_Huvec-PoisedPromoter"  
## [167] "hg19_chromatin_K562-PoisedPromoter"   
## [168] "hg19_chromatin_Nhek-PoisedPromoter"   
## [169] "hg19_chromatin_Nhlf-PoisedPromoter"   
## [170] "hg19_chromatin_Gm12878-StrongEnhancer"
## [171] "hg19_chromatin_H1hesc-StrongEnhancer" 
## [172] "hg19_chromatin_Hepg2-StrongEnhancer"  
## [173] "hg19_chromatin_Hmec-StrongEnhancer"   
## [174] "hg19_chromatin_Hsmm-StrongEnhancer"   
## [175] "hg19_chromatin_Huvec-StrongEnhancer"  
## [176] "hg19_chromatin_K562-StrongEnhancer"   
## [177] "hg19_chromatin_Nhek-StrongEnhancer"   
## [178] "hg19_chromatin_Nhlf-StrongEnhancer"   
## [179] "hg19_chromatin_Gm12878-WeakEnhancer"  
## [180] "hg19_chromatin_H1hesc-WeakEnhancer"   
## [181] "hg19_chromatin_Hepg2-WeakEnhancer"    
## [182] "hg19_chromatin_Hmec-WeakEnhancer"     
## [183] "hg19_chromatin_Hsmm-WeakEnhancer"     
## [184] "hg19_chromatin_Huvec-WeakEnhancer"    
## [185] "hg19_chromatin_K562-WeakEnhancer"     
## [186] "hg19_chromatin_Nhek-WeakEnhancer"     
## [187] "hg19_chromatin_Nhlf-WeakEnhancer"     
## [188] "hg19_chromatin_Gm12878-Insulator"     
## [189] "hg19_chromatin_H1hesc-Insulator"      
## [190] "hg19_chromatin_Hepg2-Insulator"       
## [191] "hg19_chromatin_Hmec-Insulator"        
## [192] "hg19_chromatin_Hsmm-Insulator"        
## [193] "hg19_chromatin_Huvec-Insulator"       
## [194] "hg19_chromatin_K562-Insulator"        
## [195] "hg19_chromatin_Nhek-Insulator"        
## [196] "hg19_chromatin_Nhlf-Insulator"        
## [197] "hg19_chromatin_Gm12878-TxnTransition" 
## [198] "hg19_chromatin_H1hesc-TxnTransition"  
## [199] "hg19_chromatin_Hepg2-TxnTransition"   
## [200] "hg19_chromatin_Hmec-TxnTransition"    
## [201] "hg19_chromatin_Hsmm-TxnTransition"    
## [202] "hg19_chromatin_Huvec-TxnTransition"   
## [203] "hg19_chromatin_K562-TxnTransition"    
## [204] "hg19_chromatin_Nhek-TxnTransition"    
## [205] "hg19_chromatin_Nhlf-TxnTransition"    
## [206] "hg19_chromatin_Gm12878-TxnElongation" 
## [207] "hg19_chromatin_H1hesc-TxnElongation"  
## [208] "hg19_chromatin_Hepg2-TxnElongation"   
## [209] "hg19_chromatin_Hmec-TxnElongation"    
## [210] "hg19_chromatin_Hsmm-TxnElongation"    
## [211] "hg19_chromatin_Huvec-TxnElongation"   
## [212] "hg19_chromatin_K562-TxnElongation"    
## [213] "hg19_chromatin_Nhek-TxnElongation"    
## [214] "hg19_chromatin_Nhlf-TxnElongation"    
## [215] "hg19_chromatin_Gm12878-WeakTxn"       
## [216] "hg19_chromatin_H1hesc-WeakTxn"        
## [217] "hg19_chromatin_Hepg2-WeakTxn"         
## [218] "hg19_chromatin_Hmec-WeakTxn"          
## [219] "hg19_chromatin_Hsmm-WeakTxn"          
## [220] "hg19_chromatin_Huvec-WeakTxn"         
## [221] "hg19_chromatin_K562-WeakTxn"          
## [222] "hg19_chromatin_Nhek-WeakTxn"          
## [223] "hg19_chromatin_Nhlf-WeakTxn"          
## [224] "hg19_chromatin_Gm12878-Repressed"     
## [225] "hg19_chromatin_H1hesc-Repressed"      
## [226] "hg19_chromatin_Hepg2-Repressed"       
## [227] "hg19_chromatin_Hmec-Repressed"        
## [228] "hg19_chromatin_Hsmm-Repressed"        
## [229] "hg19_chromatin_Huvec-Repressed"       
## [230] "hg19_chromatin_K562-Repressed"        
## [231] "hg19_chromatin_Nhek-Repressed"        
## [232] "hg19_chromatin_Nhlf-Repressed"        
## [233] "hg19_chromatin_Gm12878-Heterochrom/lo"
## [234] "hg19_chromatin_H1hesc-Heterochrom/lo" 
## [235] "hg19_chromatin_Hepg2-Heterochrom/lo"  
## [236] "hg19_chromatin_Hmec-Heterochrom/lo"   
## [237] "hg19_chromatin_Hsmm-Heterochrom/lo"   
## [238] "hg19_chromatin_Huvec-Heterochrom/lo"  
## [239] "hg19_chromatin_K562-Heterochrom/lo"   
## [240] "hg19_chromatin_Nhek-Heterochrom/lo"   
## [241] "hg19_chromatin_Nhlf-Heterochrom/lo"   
## [242] "hg19_chromatin_Gm12878-Repetitive/CNV"
## [243] "hg19_chromatin_H1hesc-Repetitive/CNV" 
## [244] "hg19_chromatin_Hepg2-Repetitive/CNV"  
## [245] "hg19_chromatin_Hmec-Repetitive/CNV"   
## [246] "hg19_chromatin_Hsmm-Repetitive/CNV"   
## [247] "hg19_chromatin_Huvec-Repetitive/CNV"  
## [248] "hg19_chromatin_K562-Repetitive/CNV"   
## [249] "hg19_chromatin_Nhek-Repetitive/CNV"   
## [250] "hg19_chromatin_Nhlf-Repetitive/CNV"   
## [251] "hg19_enhancers_fantom"                
## [252] "hg38_enhancers_fantom"                
## [253] "mm9_enhancers_fantom"                 
## [254] "mm10_enhancers_fantom"                
## [255] "hg19_lncrna_gencode"                  
## [256] "hg38_lncrna_gencode"                  
## [257] "mm10_lncrna_gencode"                  
## [258] "dm3_basicgenes"                       
## [259] "dm6_basicgenes"                       
## [260] "galGal5_basicgenes"                   
## [261] "hg19_basicgenes"                      
## [262] "hg38_basicgenes"                      
## [263] "mm9_basicgenes"                       
## [264] "mm10_basicgenes"                      
## [265] "rn4_basicgenes"                       
## [266] "rn5_basicgenes"                       
## [267] "rn6_basicgenes"                       
## [268] "galGal5_cpgs"                         
## [269] "hg19_cpgs"                            
## [270] "hg38_cpgs"                            
## [271] "mm9_cpgs"                             
## [272] "mm10_cpgs"                            
## [273] "rn4_cpgs"                             
## [274] "rn5_cpgs"                             
## [275] "rn6_cpgs"                             
## [276] "hg19_Gm12878-chromatin"               
## [277] "hg19_H1hesc-chromatin"                
## [278] "hg19_Hepg2-chromatin"                 
## [279] "hg19_Hmec-chromatin"                  
## [280] "hg19_Hsmm-chromatin"                  
## [281] "hg19_Huvec-chromatin"                 
## [282] "hg19_K562-chromatin"                  
## [283] "hg19_Nhek-chromatin"                  
## [284] "hg19_Nhlf-chromatin"
  1. Pathway enrichment analysis
library(TxDb.Hsapiens.UCSC.hg19.knownGene)
txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene

for (dataset in names(data)){
  genes <- seq2gene(data[[dataset]], tssRegion = c(-3000, 3000), flankDistance = 3000, TxDb=txdb)
  kkm <- enrichKEGG(gene         = genes,
                 organism     = 'human',
                 pvalueCutoff = 0.05)
  

  print(dotplot(kkm))
  
  engo <- enrichGO(gene          = genes,
                OrgDb         = org.Hs.eg.db,
                ont           = "BP",
                pAdjustMethod = "BH",
                pvalueCutoff  = 0.01,
                qvalueCutoff  = 0.05,
        readable      = TRUE)
  print(dotplot(engo))
  
  }
## Reading KEGG annotation online:
## 
## Reading KEGG annotation online: