Using CisGenome to Analyze ChIP‐chip and ChIP‐seq Data
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Abstract
Chromatin immunoprecipitation (ChIP) coupled with genome tiling array hybridization (ChIP?chip) and ChIP followed by massively parallel sequencing (ChIP?seq) are high?throughput approaches to profiling genome?wide protein?DNA interactions. Both technologies are increasingly used to study transcription?factor binding sites and chromatin modifications. CisGenome is an integrated software system for analyzing ChIP?chip and ChIP?seq data. This unit describes basic functions of CisGenome and how to use them to find genomic regions with protein?DNA interactions, visualize binding signals, associate binding regions with nearby genes, search for novel transcription?factor binding motifs, and map existing DNA sequence motifs to user?supplied genomic regions to define their exact locations.Curr. Protoc. Bioinform. 33:2.13.1?2.13.45. © 2011 by John Wiley & Sons, Inc.
Keywords: transcription factor; chromatin immunoprecipitation; tiling array; next generation sequencing; motif; gene regulation
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PDF or HTML at Wiley Online LibraryTable of Contents
- Introduction
- Basic Protocol 1: ChIP‐chip Peak Calling for Affymetrix Tiling Array Data
- Basic Protocol 2: Visualization
- Basic Protocol 3: Peak‐Gene Association
- Basic Protocol 4: DNA Sequence Retrieval
- Basic Protocol 5: De Novo Motif Discovery
- Basic Protocol 6: Motif Mapping
- Basic Protocol 7: ChIP‐chip Peak Calling for Other Tiling Array Platforms
- Basic Protocol 8: ChIP‐seq Peak Calling (One‐Sample Analysis)
- Basic Protocol 9: ChIP‐seq Peak Calling (Two‐Sample Analysis)
- Support Protocol 1: Installing CisGenome
- Support Protocol 2: Installing Genome Databases
- Guidelines for Understanding Results
- Commentary
- Literature Cited
- Figures
- Tables
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Figure 2.13.1 Overview of the CisGenome basic data analysis pipeline. View Image -
Figure 2.13.2 The CisGenome graphic user interface (GUI) and menu system. The menu for creating an Affymetrix tiling array data set is shown as an example. View Image -
Figure 2.13.3 The dialog for adding BPMAP files to an Affymetrix ChIP‐chip data set. View Image -
Figure 2.13.4 The dialog for adding CEL files to an Affymetrix ChIP‐chip data set. View Image -
Figure 2.13.5 The newly created tiling array data set shown in the CisGenome Project Explorer. Double‐clicking a CEL file will open a CisGenome Browser window displaying a heat map of the array image. View Image -
Figure 2.13.6 The dialog for normalizing an Affymetrix tiling array data set. View Image -
Figure 2.13.7 ChIP‐chip peak calling. Before peak detection, a normalized tiling array data set (circle 1.10) needs to be available in the Project Explorer, and one needs to provide several basic peak calling parameters in a dialog. View Image -
Figure 2.13.8 ChIP‐chip peak calling results. Peaks are summarized in a COD file shown in the right window. A number of BAR files are also created to store enrichment signals. Both the COD file and the BAR files are added to the Project Explorer on the left. View Image -
Figure 2.13.9 CisGenome Browser. (A ) The shortcut icon for the browser. (B ) The first page of the browser. View Image -
Figure 2.13.10 The browser page for choosing browser session type. View Image -
Figure 2.13.11 An empty browser session newly created. View Image -
Figure 2.13.12 The browser page for choosing data track type. View Image -
Figure 2.13.13 The track configuration page in CisGenome Browser. View Image -
Figure 2.13.14 CisGenome Browser showing different types of data. Tools to adjust the display styles are highlighted. View Image -
Figure 2.13.15 Peak‐gene association. (A ) The dialog for annotate peaks by nearby genes. (B ) The annotation results returned in a COD file. View Image -
Figure 2.13.16 DNA sequence retrieval. (A ) The parameter configuration dialog. (B ) Returned files. The DNA sequences will be returned in FASTA format (top). If cross‐species conservation score is requested, conservation scores for each sequence will be returned as well. The conservation scores can be returned in a text format (bottom left), in BED format (bottom right), or a binary CS format (not shown). View Image -
Figure 2.13.17 The parameter configuration dialog for de novo motif discovery. View Image -
Figure 2.13.18 An example of the summary file produced by de novo motif discovery. View Image -
Figure 2.13.19 Motif matrix files produced by de novo motif discovery. (A ) Each motif matrix is stored in a MAT file. (B ) The list of motifs is stored in a MATL file. (C ) Double‐clicking the MATL file in Project Explorer opens CisGenome Browser to display sequence logos of the motifs. The last motif in this example matches the known Gli motif. View Image -
Figure 2.13.20 An example of the CONS file for describing motif consensus sequence. View Image -
Figure 2.13.21 Mapping a motif matrix to a list of genomic regions. (A ) The parameter configuration dialog. (B ) The mapped motif sites are saved to a COD file. View Image -
Figure 2.13.22 Input data format for calling peaks from ChIP‐chip experiments based on non‐Affymetrix tiling array platforms. View Image -
Figure 2.13.23 The parameter configuration dialog for normalizing ChIP‐chip data from a text file. View Image -
Figure 2.13.24 Converting ChIP‐chip data in a text file to a tiling array data set consisting of BAR files. (A ) The parameter configuration dialog. (B ) The converted data set shown in Project Explorer. View Image -
Figure 2.13.25 A sample ALN file. View Image -
Figure 2.13.26 Loading aligned reads for ChIP‐seq peak calling. (A ) The parameter configuration dialog for loading the ALN file. (B ) Loaded data shown in Project Explorer. View Image -
Figure 2.13.27 FDR computation for an one‐sample ChIP‐seq experiment. (A ) The parameter configuration dialog. (B ) The results are returned in a table that summarizes statistical properties of the data. View Image -
Figure 2.13.28 Peak calling from one‐sample ChIP‐seq data. (A ) The parameter configuration dialog. (B ) The detected peaks are reported in a COD file. View Image -
Figure 2.13.29 Data for two‐sample ChIP‐seq analysis loaded into CisGenome. View Image -
Figure 2.13.30 FDR computation for a two‐sample ChIP‐seq experiment. (A ) The parameter configuration dialog. (B ) The results are returned in a table that summarizes statistical properties of the data. View Image -
Figure 2.13.31 The parameter configuration dialog for two‐sample ChIP‐seq peak calling. View Image -
Figure 2.13.32 An example of the CisGenome.ini file. View Image -
Figure 2.13.33 Load a genome database into CisGenome GUI. (A ) In the file open dialog, choose the file named [species]_[assembly].cgw in the genome database folder. (B ) The loaded database shown in Project Explorer. View Image
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