ChIPpeakAnno overlap peaks with TSS returns more than TSS overlap
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94133 • 0
@94133-14305
Last seen 4.4 years ago
USA, Stanford

I want ChIP peaks that overlap gene TSSs. However, output from ChIPpeakAnno returns peaks that do not overlap, which requires extra filtering. Is there a better way?

ChIP_peaks_annoTSS <- annotatePeakInBatch(res1_ChIP,
                                            AnnotationData = genes(TxDb.Mmusculus.UCSC.mm10.knownGene),
                                            output = "overlapping",
                                            featureType = "TSS",
                                            select = "all",
                                            ignore.strand = TRUE,
                                            FeatureLocForDistance = "TSS")
ChIP_peaks_annoTSS <- addGeneIDs(annotatedPeak=ChIP_peaks_annoTSS,
                                   orgAnn = "org.Mm.eg.db",
                                   feature_id_type = "entrez_id",
                                   IDs2Add = "symbol") %>% as.data.frame()

fromOverlappingOrNearest column = Overlapping, when insideFeature shows inside or overlapEnd, which is NOT TSS.

So then, I filter from insideFeature column to get TSS overlaps, like:

TSSpatterns = c("overlapStart","includeFeature")
ChIP_peaks_annoTSS <- filter(ChIP_peaks_annoTSS, grepl(paste(TSSpatterns, collapse="|"), insideFeature))
ChIP_peaks_annoTSS_cond <- condenseMatrixByColnames(as.matrix(as.data.frame(ChIP_peaks_annoTSS)), "peak")

Can you show me the proper way?

Thanks!!!!

 

 

chippeakanno chipseq R • 1.8k views
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Could you please try the following code and see if that meets your need? Thanks!

tss <- promoters(TxDb.Mmusculus.UCSC.mm10.knownGene, upstream=0, downstream=1)

ChIP_peaks_annoTSS <- annotatePeakInBatch(res1_ChIP,
                                            AnnotationData = tss,
                                            output = "overlapping",
                                            featureType = "TSS",
                                            select = "all",
                                            ignore.strand = TRUE,
                                            FeatureLocForDistance = "TSS")

Best regards,

Julie

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Ou, Jianhong ★ 1.3k
@ou-jianhong-4539
Last seen 2 days ago
United States

Did you tried to set output = "upstream"?

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No. Are you suggesting this is the best way to do this? I don't understand why one would use upstream for TSS overlap, can you explain?

Thanks!

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This will find the peaks overlap with the TSS because we set the maxgap=-1 and FeatureLocForDistance="TSS". 

However, maybe this is not the answer of your biological question. Maybe you are asking to find the annotation for promoter region? If that is the case, please try to use set output="overlapping", FeatureLocForDistance="TSS" and bindingRegion = c(-5000, 3000). Here the bindingRegion means upstream 5K and downstream 3K of TSS.

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I tried your suggestion like this but get an error:

ChIP_peaks_annoTSS <- annotatePeakInBatch(res1_ChIP,

                                            AnnotationData = genes(TxDb.Mmusculus.UCSC.mm10.knownGene),
                                            output = "overlapping",
                                            featureType = "TSS",
                                            select = "all",
                                            ignore.strand = TRUE,
                                            FeatureLocForDistance = "TSS",
                                            bindingRegion = c(-2000, 2000))
ChIP_peaks_annoTSS <- addGeneIDs(annotatedPeak=ChIP_peaks_annoTSS,
                                   orgAnn = "org.Mm.eg.db",
                                   feature_id_type = "entrez_id",
                                   IDs2Add = "symbol") 
ChIP_peaks_annoTSS_cond <- condenseMatrixByColnames(as.matrix(as.data.frame(ChIP_peaks_annoTSS)), "peak")

Error in data.frame(seqnames = as.factor(seqnames(x)), start = start(x),  : 
  duplicate row.names: X12, X39, X45, X52, X67, X71, X137, X144, X179, X184, X215, X228, X232, X240, X244, X246, X255, X262, X265, X284, X287, X379, X384, X391, X393, X404, X420, X451, X533, X534, X536, X553, X556, X574, X575, X60 ... ... ... 

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try:

ChIP_peaks_annoTSS_cond <- condenseMatrixByColnames(as.matrix(as.data.frame(unname(ChIP_peaks_annoTSS))), "peak")

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That works, thanks! 

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