Gokegg Analysis

Source: https://github.com/aipoch/medical-research-skills

When To Read External Files

Situation	File To Read	Purpose
Need algorithm details	references/algorithm.md	Statistical methods and formulas
Need to run the analysis	scripts/main.R	Full execution command
Encounter an error	references/troubleshooting.md	Troubleshooting guidance
Need CLI examples	references/cli-guide.md	Parameter usage examples

When To Use

Use this skill for:

• GO and KEGG enrichment from a gene list derived from bulk RNA-seq or microarray studies
• Supported gene ID types: SYMBOL, ENSEMBL, ENTREZID
• Supported species databases: org.Hs.eg.db, org.Mm.eg.db, org.Rn.eg.db

Do not use this skill for:

• Single-cell RNA-seq analysis
• Methylation, proteomics, or non-expression omics workflows
• Differential expression testing from raw count matrices

Usage

Main analysis and plotting: Rscript scripts/main.R --feature "TP53,EGFR,BRCA1,MYC" --output_dir ./output --sp org.Hs.eg.db --gene_type SYMBOL --pvalue_cutoff 0.05 --qvalue_cutoff 0.2 --pAdjustMethod BH --seed 66 --go_top_n 3 --kegg_top_n 3 --format pdf

Notes:

• scripts/main.R is the only command-line entry point
• scripts/dochart.R currently provides plotting functions and is sourced by scripts/main.R
• If --go_input, --kegg_input, or --outdir are omitted, main.R uses output_dir/temp/GO_list.rda, output_dir/temp/KEGG_list.rda, and output_dir/plot automatically

Agent Output

On success, the agent should report:

• Whether GO enrichment completed successfully
• Whether KEGG enrichment completed successfully
• The normalized input gene count after trimming and parsing
• The main output directory
• The generated files, especially GO_df.csv, KEGG_df.csv, GO_list.rda, KEGG_list.rda, and the combined dot chart
• The path to session_info.txt

Post-run checklist:

• Re-parse the original --feature string using the documented separator rules and report the deduplicated gene count after trimming
• Check temp/GO_df.csv and temp/GO_list.rda before claiming GO success
• Check temp/KEGG_df.csv and temp/KEGG_list.rda before claiming KEGG success
• Check plot/gokegg_dot_chart.<format>, plot/gokegg_dot_chart_data.csv, plot/gokegg_dot_chart_data.rda, and session_info.txt before claiming full success
• Summarize the final result with: parsed gene count, GO status, KEGG status, plot status, output directory, and key output files

On failure, the agent should report:

• The exact SKILL_* error code
• The failing step, such as gene parsing, ID conversion, enrichment, or plotting
• The actionable next step, such as fixing input IDs, checking missing packages, or regenerating .rda files

Parameter Reference

scripts/main.R

Short	Long	Type	Default	Required	Description
-f	--feature	character	""	Yes	Gene list separated by commas, Chinese commas, semicolons, tabs, or newlines
-o	--output_dir	character	./output/	No	Main output directory
-s	--sp	character	org.Hs.eg.db	No	Species database
-g	--gene_type	character	SYMBOL	No	Input gene ID type
-p	--pvalue_cutoff	numeric	0.05	No	Enrichment p-value cutoff
-q	--qvalue_cutoff	numeric	0.2	No	Enrichment q-value cutoff
-m	--pAdjustMethod	character	BH	No	P-value adjustment method
	--seed	integer	66	No	Random seed
	--go_input	character	NULL	No	Optional GO .rda; defaults to output_dir/temp/GO_list.rda
	--kegg_input	character	NULL	No	Optional KEGG .rda; defaults to output_dir/temp/KEGG_list.rda
	--outdir	character	NULL	No	Plot output directory; defaults to output_dir/plot
	--go_top_n	numeric	3	No	Top GO terms per ontology
	--kegg_top_n	numeric	3	No	Top KEGG pathways
-w	--width	numeric	20	No	Plot width in cm
	--height	numeric	16	No	Plot height in cm
	--format	character	pdf	No	Plot format: pdf, png, svg
	--dpi	numeric	300	No	DPI for raster output
-c	--colors	character	#E41A1C,#FFFF33,#2E86AB,#4DAF4A	No	Colors for GO:BP,GO:CC,GO:MF,KEGG
	--title	character	GO + KEGG Dot Chart	No	Plot title
	--xlab	character	NULL	No	Horizontal axis label override
	--ylab	character	NULL	No	Vertical axis label override
	--dot_size	numeric	4.5	No	Dot size
	--shape	numeric	19	No	Dot shape
	--rotate / --no-rotate	logical flag	TRUE	No	Rotate plot orientation on or off
	--sorting	character	descending	No	Dot sorting order
	--label_width	numeric	35	No	Label wrap width
	--title_size	numeric	12	No	Title font size
	--axis_title_size	numeric	9	No	Axis title font size
	--axis_text_size	numeric	8	No	Axis text font size
	--legend_title_size	numeric	8	No	Legend title font size
	--legend_text_size	numeric	7	No	Legend text font size
	--legend_position	character	top	No	Legend position
	--plot_margin	character	10,10,10,10	No	Plot margins: top,right,bottom,left
	--axis_line_size	numeric	0.5	No	Axis line width
	--axis_ticks_size	numeric	0.5	No	Axis tick width
	--show_grid	logical	FALSE	No	Show grid lines
-v	--verbose	logical	FALSE	No	Enable verbose logging

Input Format

Main Analysis Input

• --feature should be provided as a gene list
• Preferred separator: comma
• Also accepted: Chinese commas, semicolons, tabs, and newlines
• Leading and trailing spaces around each gene are removed automatically with trimming
• The gene ID type must match --gene_type
• --sp supports only org.Hs.eg.db, org.Mm.eg.db, and org.Rn.eg.db

Examples: TP53,EGFR,BRCA1,MYC

TP53, EGFR, BRCA1, MYC

TP53；EGFR；BRCA1；MYC

TP53\nEGFR\nBRCA1\nMYC

Example command with minimal input: Rscript scripts/main.R --feature "TP53,EGFR,BRCA1,MYC" --output_dir ./example_output --sp org.Hs.eg.db --gene_type SYMBOL

Example command with custom plotting parameters: Rscript scripts/main.R --feature "TP53,EGFR,BRCA1,MYC" --output_dir ./example_plot_output --sp org.Hs.eg.db --gene_type SYMBOL --go_top_n 5 --kegg_top_n 8 --colors "#E41A1C,#FFFF33,#2E86AB,#4DAF4A" --title "Custom GO + KEGG Dot Chart" --xlab="-log10(adjusted p-value)" --ylab="Enriched Terms" --width 24 --height 18 --label_width 40 --format png --dpi 300 --no-rotate --verbose

Note: values passed to --xlab or --ylab that start with - should use --option=value syntax to avoid being parsed as flags.

Note: separator variants are supported only when they are passed inside a single --feature argument value.

Plot Input

• Plotting is triggered by scripts/main.R
• --go_input: optional .rda file containing a GO_list object
• --kegg_input: optional .rda file containing a KEGG_list object
• If not provided, main.R uses the newly generated files under output_dir/temp
• Plotting requires result tables with at least Description and p.adjust

Output Files

File Name	Format	Description
temp/GO_df.csv	CSV	GO enrichment result table
temp/GO_list.rda	RDA	Full GO enrichment object
temp/KEGG_df.csv	CSV	KEGG enrichment result table
temp/KEGG_list.rda	RDA	Full KEGG enrichment object
plot/gokegg_dot_chart.pdf etc.	PDF/PNG/SVG	Combined GO/KEGG dot chart
plot/gokegg_dot_chart_data.csv	CSV	Combined plotting table used for the figure
plot/gokegg_dot_chart_data.rda	RDA	Plot bundle with plotting data and parameters
session_info.txt	TXT	Runtime session information

Error Handling

Common error codes and fixes:

• SKILL_FILE_NOT_FOUND: Input file does not exist; check the path and permissions
• SKILL_FILE_FORMAT_ERROR: .rda cannot be read or is malformed; regenerate upstream results
• SKILL_MISSING_COLUMNS: Result table is missing Description or p.adjust
• SKILL_EMPTY_DATA: Input genes are empty after parsing, cannot be converted, or enrichment results are empty
• SKILL_INVALID_PARAMETER: Required parameter missing, unsupported species, or insufficient color count
• SKILL_PACKAGE_NOT_FOUND: Required package is not installed
• SKILL_ANALYSIS_FAILED: Internal GO/KEGG enrichment failure; verify gene_type, sp, and input genes

For detailed troubleshooting, read references/troubleshooting.md.

Testing

Minimal test dataset: use a small built-in gene list directly, with no extra files required.

Smoke test command: Rscript scripts/main.R --feature "TP53,EGFR,BRCA1,MYC" --output_dir ./test_output --sp org.Hs.eg.db --gene_type SYMBOL --pvalue_cutoff 0.05 --qvalue_cutoff 0.2 --pAdjustMethod BH --seed 66 --go_top_n 3 --kegg_top_n 3 --format pdf --verbose

Expected smoke-test outputs:

• ./test_output/temp/GO_list.rda
• ./test_output/temp/KEGG_list.rda
• ./test_output/temp/GO_df.csv
• ./test_output/temp/KEGG_df.csv
• ./test_output/plot/gokegg_dot_chart.pdf
• ./test_output/plot/gokegg_dot_chart_data.csv
• ./test_output/plot/gokegg_dot_chart_data.rda
• ./test_output/session_info.txt
• Exit status code 0

Automated regression script: Rscript test/test_regressions.R

The regression script covers:

• Separator parsing with comma, Chinese semicolon, newline, tab, and mixed separators
• Empty parsed-gene handling
• Invalid --plot_margin validation
• Plot input validation for missing GO/KEGG inputs

Separator examples for manual CLI verification: Rscript scripts/main.R --feature "TP53,EGFR,BRCA1,MYC" --output_dir ./test_sep_comma Rscript scripts/main.R --feature "TP53；EGFR；BRCA1；MYC" --output_dir ./test_sep_cn_semicolon Rscript scripts/main.R --feature $'TP53\nEGFR\nBRCA1\nMYC' --output_dir ./test_sep_newline Rscript scripts/main.R --feature $'TP53\tEGFR\tBRCA1\tMYC' --output_dir ./test_sep_tab Rscript scripts/main.R --feature $'TP53； EGFR, BRCA1 MYC' --output_dir ./test_sep_mixed

Note: all separators must be passed inside a single --feature argument value.