Here is a concise summary of the key steps and tools for ChIP-seq and RNA-seq data analysis and integration:
-
Quality control: FastQC for assessing raw sequencing data quality.
-
Trimming and filtering: Trimmomatic or Cutadapt for preprocessing reads.
-
Alignment: Bowtie2 or BWA for ChIP-seq, and STAR, HISAT2, or TopHat2 for RNA-seq.
-
Peak calling (ChIP-seq): MACS2, SICER, HOMER (see separate article) or diffReps.pl (part of the DiffReps package) for identifying bound genomic regions.
-
Gene expression quantification (RNA-seq): featureCounts, HTSeq, or Cufflinks for expression levels.
-
Differential expression analysis (RNA-seq): DESeq2 or edgeR for comparing conditions or time points.
-
Motif analysis (ChIP-seq): MEME-ChIP for identifying enriched sequence motifs.
-
Data visualization: deepTools or Integrative Genomics Viewer (IGV) for viewing aligned reads and peaks.
-
Annotation and integration: ChIPseeker or HOMER for peak annotation, and GenomicRanges, DiffBind, or other R packages for integrating ChIP-seq and RNA-seq data.
-
Functional enrichment analysis: GSEA, clusterProfiler, or DAVID for pathway and functional category enrichment.
-
Visualization: ggplot2 or ComplexHeatmap for combined ChIP-seq and RNA-seq data.