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2022-01-17 cellmarker注释

2022-01-17 cellmarker注释

作者: 徐添添 | 来源:发表于2022-01-18 00:02 被阅读0次

    细胞类型自动化注释

    # 数据来源:http://biocc.hrbmu.edu.cn/CellMarker/download.jsp
    library(dplyr)
    library(openxlsx)
    #### 1. 加载marker数据库 ####
    load("./Human_cell_markers.RData")
    cellmarker.file <- "./multi/multi_total_marker_genes_tsne_20PC.txt"
    sampleid <- "multi"
    #读入单细胞分析中输出的cell marker 基因文件
    marker.gene <- read.table(cellmarker.file,header = T,stringsAsFactors = F,sep = "\t")
    marker.gene.sig <- marker.gene %>% filter(as.numeric(p_val_adj) <= 0.05)
    
    cat("Totally marker genes:",length(unique(marker.gene.sig$gene)))
    table(marker.gene.sig$cluster)
    
    #### 2. 定位数据库中存在的基因 ####
    marker.gene.sig %>% filter(gene %in% cell.markers.tb$geneSymbol) -> marker.gene.sel
    marker.gene.sel$cellMarker <- apply(marker.gene.sel,1,function(x)paste(unique(cell.markers.tb[cell.markers.tb$geneSymbol == x["gene"],1]),collapse = ","))
    cat("Totally find",length(unique(marker.gene.sel$gene)),"/",length(unique(marker.gene.sig$gene)),"genes in cellMarker db")
    
    #### 3. 将marker基因与细胞类型结果写出到文件 ####
    write.table(marker.gene.sel,file =paste0("./",sampleid,"/",sampleid,"_DEG_marker_cells_tsne.txt"),row.names = T,col.names = T,sep = "\t",quote = F)
    #sub_analysis_scRNA
    

    以下代码在sub_analysis_scRNA.R中
    计算特定两组细胞之间的差异基因,决定要不要把两个亚群合并

    # 计算特定两组细胞之间的差异基因
    sub.markers <- FindMarkers(experiment.aggregate,
                               ident.1 = c("0","3"),
                               ident.2 = "1")
    View(sub.markers)
    

    修改具体类别名字

    # 修改具体类别的名字(需要基于marker基因来定义)
    experiment.merged <- RenameIdents(
      object = experiment.aggregate,
      "0" = "B cells type1",
      "2" = "B cells type2",
      "3" = "B cells type1"
    )
    pdf(paste0("./",sam.name,"/CellCluster-TSNEPlot_Rename_",max(dim.use),"PC.pdf"),width = 5,height = 4)
    DimPlot(object = experiment.merged, pt.size=0.5,
            reduction = "tsne",label = T) +
      ggsci::scale_color_igv()
    dev.off()
    

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