pseudobulkdeg

Aggregate single-cell counts into pseudo-bulk samples and test differential expression with DESeq2 or edgeR.

22|4|Updated May 18, 2021
One-click install
npx skills add https://github.com/pwwang/immunopipe --skill pseudobulkdeg
Or copy as Structured Prompt for Agent▼
Please help me install this Agent Skill.
Skill: pseudobulkdeg
Source: https://github.com/pwwang/immunopipe/tree/main/skills/pseudobulkdeg
Command: npx skills add https://github.com/pwwang/immunopipe --skill pseudobulkdeg

SYSTEM DOCUMENTATION & REQUIREMENTS

What problem does it solve?

Aggregates single-cell counts into sample-level pseudo-bulk data and identifies differentially expressed genes between conditions, accounting for biological replicates.

Core Features & Use Cases

  • Supports complex experimental designs including batch effects, paired samples, and interactions.
  • Enables DE analysis with DESeq2 or edgeR and automatically performs enrichment on significant markers.
  • Applies per-cell-type analysis and produces publication-ready results across multiple conditions.

Quick Start

Provide a minimal configuration to compare two conditions after aggregating cells into pseudo-bulk samples.

Frequently Asked Questions about pseudobulkdeg

High-intent search queries and answers about installing and using this skill.

FAQPage Schema
How do I perform pseudobulk differential expression analysis on scRNA-seq data?▼

You can perform pseudobulk DEG analysis on scRNA-seq data by aggregating single-cell counts into sample-level data and testing differential expression with DESeq2 or edgeR. This approach correctly accounts for biological replicates and per-cell-type variations across conditions.

Can I use pseudobulk DEG analysis for experimental designs with batch effects and paired samples?▼

Yes, pseudobulk DEG analysis supports complex experimental designs including batch effects, paired samples, and interactions across multiple conditions. Aggregating single-cell counts into sample-level pseudo-bulk data allows standard bulk RNA-seq models to account for these confounding factors robustly.

What is the difference between using DESeq2 and edgeR for scRNA-seq pseudobulk analysis?▼

DESeq2 and edgeR are both supported for pseudobulk differential expression testing. The choice between them depends on your preferred statistical modeling framework, while the aggregation step consistently groups single-cell counts into sample-level matrices before either tool performs the testing and optional enrichment.

How do I run per-cell-type differential expression across multiple conditions?▼

To run per-cell-type differential expression, the workflow groups metadata by cell type and aggregates single-cell counts into sample-level pseudo-bulk profiles. It then performs differential expression testing across multiple conditions and automatically generates publication-ready results and optional enrichment plots.

Does pseudobulk DEG analysis automatically perform enrichment on significant markers?▼

Yes, pseudobulk DEG analysis automatically performs enrichment analysis on significant differentially expressed markers. After aggregating scRNA-seq counts and testing with DESeq2 or edgeR, the workflow includes optional enrichment and plotting steps to produce publication-ready results.