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##Phaedra Agius, April 2019
##
##INPUT: scn=urls to scn files output by SCANVIS scan or linkvar functions
## OR scn can also be a list of matrices
## method='mean' or 'median'
## roi=region of interest chr,start,end
## gen=gencode data
##
##OUTPUT: object with components sj, sj.samples and muts (muts available only if
## scn are outputs of SCANVISlinkvar)
## out$sj=averaged RRS and uniq.reads across samples
## out$sj.samples=comma sep samples, RRSs and uniq.reads for each sj
## out$muts=all mutations mapped to sjs in out$sj, with comma sep
## samples and number of samples containing the mutation
SCANVISmerge<-function(scn,method='mean',roi=NULL,gen=NULL){
roi.g=NULL
if(length(roi)>0){
if(length(grep('chr',roi))==0){
if(length(gen)==0){
stop('Please supply gencode details so that the
appropriate genomic region for your query genes can be derived')
}
roi.g=roi
roi=gene2roi(roi,gen)
}
}
if(length(roi)==0 & length(scn)>50){
stop('For more than 50 samples, we recommend merging or agglomerating
samples for a query region or chr,
otherwise resulting data matrices will be too large')
}
#load up data
usjcc=c('chr','start','end','JuncType','gene_name','FrameStatus')
roi.nn=NULL
if(length(roi)>1)
roi.nn=as.numeric(roi[2:3])
if(is.list(scn)){
sj=scn
names(sj)=names(scn)
if(length(roi)>0)
sj=lapply(sj,function(x) x[which(x[,'chr']==roi[1]),])
if(length(roi)>1){
sj=lapply(sj,function(x)
x[which(as.numeric(x[,'start'])>=roi.nn[1]),])
sj=lapply(sj,function(x)
x[which(as.numeric(x[,'end'])<=roi.nn[2]),])
}
}
if(is.character(scn)){
sj=NULL
for(i in seq(1,length(scn),1)){
id=scn[i]
tmp=gsub(' ','',as.matrix(read.delim(scn[i])))
if(length(roi)>0)
tmp=tmp[which(tmp[,'chr']==roi[1]),]
if(length(roi.nn)>0){
tmp=tmp[which(as.numeric(tmp[,'start'])>=roi.nn[1]),]
tmp=tmp[which(as.numeric(tmp[,'end'])<=roi.nn[2]),]
}
sj[[i]]=tmp
}
}
#adding rownames
for(i in seq(1,length(sj),1)){
tmp=sj[[i]][,usjcc]
h=which(tmp=='')
if(length(h)>0) tmp[h]='NA'
tmp=gsub(' ','',tmp)
rownames(sj[[i]])=apply(tmp,1,function(x) paste(x,collapse=' '))
}
#agglomerate data into matrices
usj=unique(unlist(lapply(sj,function(x) rownames(x))))
N=length(usj)
RRS=matrix(0,N,length(scn))
rownames(RRS)=usj
colnames(RRS)=names(sj)
NR=RRS
#initializing MUTS matrix
MUTS=NULL
n1=unlist(lapply(sj,function(x) ncol(x)))
n2=unlist(lapply(sj,function(x)
max(which(is.element(colnames(x),c('FrameStatus','covRRS'))))))
h=which((n1-n2)>0)
if(length(h)>0){
mm=lapply(h,function(x) sj[[x]][,(n2[x]+1):n1[x]])
tmp=unique(unlist(strsplit(unlist(mm),'\\|')))
tmp=setdiff(tmp,'')
tmp=tmp[which(!is.na(tmp))]
MUTS=matrix(0,length(tmp),length(scn))
rownames(MUTS)=tmp
colnames(MUTS)=colnames(RRS)
}
for(i in seq(1,length(scn),1)){
sj.tmp=sj[[i]]
xx=intersect(rownames(RRS),rownames(sj.tmp))
RRS[xx,i]=as.numeric(sj.tmp[xx,'RRS'])
NR[xx,i]=as.numeric(sj.tmp[xx,'uniq.reads'])
if(length(MUTS)>0){
q=max(which(is.element(colnames(sj.tmp),c('FrameStatus','covRRS'))))
if(ncol(sj.tmp)>q){
Q=(q+1):ncol(sj.tmp)
mut.new=as.vector(sj.tmp[,Q])
mut.new=mut.new[which(mut.new!='')]
mut.new=mut.new[which(!is.na(mut.new))]
mut.new=setdiff(unique(unlist(strsplit(mut.new,'\\|'))),'')
MUTS[mut.new,i]=1
}
}
}
##representative sample
S0=t(matrix(unlist(strsplit(rownames(RRS),' ')),6,nrow(RRS)))
if(method=='mean') S0=cbind(S0,apply(RRS,1,mean),apply(NR,1,mean))
if(method=='median') S0=cbind(S0,apply(RRS,1,median),apply(NR,1,median))
colnames(S0)=c('chr','start','end','JuncType',
'gene_name','FrameStatus','RRS','uniq.reads')
out=NULL
if(length(roi)>0) out$roi=roi
if(length(roi.g)>0) out$roi=c(roi,paste(roi.g,collapse=','))
out$RRS=RRS
out$NR=NR
if(length(MUTS)>0) out$MUTS=MUTS
out$SJ=S0
rownames(out$SJ)=NULL
return(out)
}
##get genomic coordinates for gene name
gene2roi<-function(g,gen){
q=which(is.element(gen$GENES[,'gene_name'],g))
tmp=intersect(g,gen$GENES[q,'gene_name'])
if(length(tmp)==0){
stop('Gene/s not found in the gencode object.
Please check your gene list')
}
if(length(tmp)!=length(g)){
print(paste('Gene/s',setdiff(g,tmp),'are not listed in the
supplied gencode object and will be excluded.'))
}
if(length(unique(gen$GENES[q,'chr']))!=1){
print(paste('The genes supplied fall into the following
chromosomes:',paste(unique(gen$GENES[q,'chr']),collapse=',')))
stop('Only one chromosome at a time is accepted.
Please supply genes in the same chromosome')
}
roi=NULL
tt=sort(table(gen$GENES[q,'chr']))
q=intersect(q,which(gen$GENES[,'chr']==tail(names(tt),1)))
if(length(unique(gen$GENES[q,'chr']))!=1){
stop('Gene list supplied occur in more than one chromosome.
Please supply gene/s that occur in one chromosome only')
}
roi=range(as.numeric(as.vector(gen$GENES[q,c('start','end')])))
roi=c(tail(names(tt)[1]),roi)
return(roi)
}
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