| name | De-novo-motif-discovery |
| description | This skill identifies novel transcription factor binding motifs in the promoter regions of genes, or directly from genomic regions of interest such as ChIP-seq peaks, ATAC-seq accessible sites, or differentially acessible regions. It employs HOMER (Hypergeometric Optimization of Motif Enrichment) to detect both known and previously uncharacterized sequence motifs enriched within the supplied genomic intervals. Use the skill when you need to uncover sequence motifs enriched or want to know which TFs might regulate the target regions. |
HOMER De Novo Motif Discovery
Overview
This skill enables comprehensive de novo motif discovery using HOMER tools for genomic peak files. It discovers novel transcription factor binding motifs from genomic regions without requiring prior knowledge of motif patterns. To perform de novo motif discovery:
- Always refer to the Inputs & Outputs section to check inputs and build the output architecture.
- Genome assembly: Always returned from user feedback (hg38, mm10, hg19, mm9, etc), never determined by yourself.
- Check chromosome names: Standardize chromosome names to format with "chr" (1 -> chr1, MT -> chrM).
- Set analysis parameters: Region size, number of motifs, motif lengths
- Run HOMER de novo motif discovery command
When to use this skill
Use this skill when you need to uncover sequence motifs enriched in the promoter regions of a set of genes, or directly from a set of genomic regions, such as peaks from ChIP-seq or ATAC-seq, without prior assumptions about which transcription factors are involved. Typical use cases include:
- Performing motif enrichment analysis in promoters of a gene list provided by user or generated in previous analysis to infer potential transcription factors that might regulate the target genes.
- Performing motif enrichment analysis in TF-binding sites or differential TF-binding regions provided by user or generated in previous analysis to infer potential transcription factors that might be co-factors of the target TFs.
- Performing motif enrichment analysis on ATAC-seq peaks or differential accessible regions provided by user or generated in previous analysis to infer potential transcriptional regulators of accessible chromatin regions.
- Exploring novel sequence patterns for the binding motif of a specific TF.
Inputs & Outputs
Inputs
Input files should be in one of the following formats:
- BED files: Standard genomic interval format
- narrowPeak: narrow peak format
- broadPeak: broad peak format
- gene list: A list of genes provided by user or generated in previous analysis. May end with .txt, .tsv, .csv, etc.
Outputs
${sample}_de_novo_motif_discovery/
results/
homerResults.html # De novo motif discovery results
seq.autonorm.tsv # Sequence composition statistics
motifFindingParameters.txt # Parameters used for analysis
homerMotifs.all.motifs
homerMotifs.motifs12
homerMotifs.motifs10
homerMotifs.motifs8
nonRedundant.motifs
homerResults/
motif1.similar1.motif
motif1.info.html
motif1.logo.svg
motif1.motif
motif1.similar.html
motif1.similar2.motif
motif1.similar3.motif
motif1.similar4.motif
motif1RV.logo.svg
motif1RV.motif
# ...
logs/ # analysis logs
motif.log
Decision Tree
Step 0 — Gather Required Information from the User
Before calling any tool, ask the user:
- Sample name (
sample): used as prefix and for the output directory${sample}_de_novo_motif_discovery. - Genome assembly (
genome): e.g.hg38,mm10,danRer11.- Never guess or auto-detect.
Step 1: Initialize Project
- Make director for this project:
Call:
mcp__project-init-tools__project_init
with:
sample: the user-provided sample nametask: de_novo_motif_discovery
The tool will:
- Create
${sample}_de_novo_motif_discoverydirectory. - Get the full path of the
${sample}_de_novo_motif_discoverydirectory, which will be used as${proj_dir}.
Step 2: Prepare genome file for homer
Call:
mcp__homer-tools__check_genome_installation
With:
genome: the user-provided genome assembly, e.g.hg38,mm10,danRer11
The tool will:
- Check if the genome is installed in HOMER.
- If not, install the genome.
Step 3 (Optional): Standardize chromosome names for BED files
This step is optional. Only perform this step if the input file is a BED file. If the input file is a gene list, skip this step.
From 1 format to chr1 format
From MT format to chrM format
Call:
mcp__file-format-tools__standardize_bed_chrom_names
with:
input_bed: the user-provided BED fileoutput_bed: the path to save the standardized BED file
The tool will:
- Standardize the chromosome names in the BED file.
- Return the path of the standardized BED file.
Step 4: De Novo Motif Discovery
Here are three options for different situations. Pick one of them based on the user's request.
- De novo + known motifs
- De novo + known motifs + background
- De novo only
Option 1: De novo + known motifs
Call:
mcp__homer-tools__find_motifs
With:
sample: the user-provided sample nameproj_dir: directory to save the de novo motif discovery results. In this skill, it is the full path of the${sample}_de_novo_motif_discoverydirectory returned bymcp__project-init-tools__project_initinput_file: the user-provided file containing genome regions or gene list. May end with.bed,.narrowPeak,.broadPeak,.txt,.tsv,.csv, etc.genome: the user-provided genome assembly, e.g.hg38,mm10,danRer11size: region size for motif finding for genome regions (default: 200). If the input file is a gene list, set to None.mask: mask repeat regions (default: True)threads: number of processors to use (default: 4)num_motifs: number of motifs to find (default: 25)lengths: motif lengths to search (default: 8,10,12)
The tool will:
- Discover motifs in the genome regions in the bed file or the promoters of the genes in the gene list. The motifs could be known motifs or de novo motifs.
- Return the path of the de novo motif discovery results under
${proj_dir}/results/directory.
Option 2: De novo + known motifs + background
Call:
mcp__homer-tools__find_motifs
With:
sample: the user-provided sample nameproj_dir: directory to save the de novo motif discovery results. In this skill, it is the full path of the${sample}_de_novo_motif_discoverydirectory returned bymcp__project-init-tools__project_initinput_file: the user-provided file containing genome regions or gene list. May end with.bed,.narrowPeak,.broadPeak,.txt,.tsv,.csv, etc.genome: the user-provided genome assembly, e.g.hg38,mm10,danRer11background_file: the user-provided file containing background genome regions or gene list. May end with.bed,.narrowPeak,.broadPeak,.txt,.tsv,.csv, etc.size: region size for motif finding for genome regions (default: 200). If the input file is a gene list, set to None.mask: mask repeat regions (default: True)threads: number of processors to use (default: 4)num_motifs: number of motifs to find (default: 25)lengths: motif lengths to search (default: 8,10,12)
The tool will:
- Discover motifs in the genome regions in the bed file or the promoters of the genes in the gene list with background genome regions or gene list provided. The motifs could be known motifs or de novo motifs.
- Return the path of the de novo motif discovery results under
${proj_dir}/results/directory.
Option 3: De novo only
Call:
mcp__homer-tools__find_motifs
With:
sample: the user-provided sample nameproj_dir: directory to save the de novo motif discovery results. In this skill, it is the full path of the${sample}_de_novo_motif_discoverydirectory returned bymcp__project-init-tools__project_initinput_file: the user-provided file containing genome regions or gene list. May end with.bed,.narrowPeak,.broadPeak,.txt,.tsv,.csv, etc.genome: the user-provided genome assembly, e.g.hg38,mm10,danRer11size: region size for motif finding for genome regions (default: 200). If the input file is a gene list, set to None.mask: mask repeat regions (default: True)threads: number of processors to use (default: 4)num_motifs: number of motifs to find (default: 25)lengths: motif lengths to search (default: 8,10,12)noknown:Trueto not use known motifs
The tool will:
- Discover motifs in the genome regions in the bed file or the promoters of the genes in the gene list without searching for known motif enrichment.
- Return the path of the de novo motif discovery results under
${proj_dir}/results/directory.
Here are additional parameters for calling mcp__homer-tools__find_motifs tool, which are not commonly used. Add these parameters only when necessary:
cpg: Enrich for CpG islands (default: False)chopify: Chop sequences into smaller fragments (default: False)norevopp: Don't search reverse complement (default: False)rna: For RNA motif finding (default: False)bits: Set information content threshold (default: None)
Quality Control and Best Practices
Pre-processing Steps
- Filter peaks: Remove low-quality or artifact peaks
- Size selection: Use appropriate region size (-size parameter)
- Background selection: Choose appropriate background for enrichment analysis
- Repeat masking: Use
-maskfor cleaner motif discovery
Parameter Optimization
- Region size: Typically 200-500bp for transcription factors
- Motif length: 8-12bp for most transcription factors
- Number of motifs: 10-25 for initial discovery
- Threads: Use available CPU cores for faster processing
Troubleshooting
Common Issues
- Memory errors: Reduce region size or number of motifs
- Slow performance: Use
-poption for parallel processing - No motifs found: Check input file format and region size
- Genome not found: Verify genome assembly name and installation
Error Handling
- Ensure HOMER is properly installed and configured
- Check that genome data is downloaded and accessible
- Verify input file formats and chromosome naming
- Ensure sufficient disk space for output files