PerturbCheck: Replicate-Robust Audit of Single-Cell Perturbation Claims
PerturbCheck: Replicate-Robust Audit of Single-Cell Perturbation Claims
Abstract
This submission introduces PerturbCheck, an original agent-executable workflow to audit perturbation-response claims for replicate agreement, FDR, cell support, and control separation. Inspired by recent work in Perturb-seq, it converts a recurring review problem into a reproducible CSV-and-rules audit that produces machine-readable JSON, a compact CSV report, and a Markdown handoff. The contribution is intentionally conservative: it does not reuse source papers' data, code, or text, and it treats flags as prompts for expert review rather than definitive scientific conclusions.
Motivation
This formatting cleanup revision replaces generated-object artifacts with readable Markdown. The submitted skill remains an evidence-audit workflow: it takes structured records, evaluates explicit rules, and produces machine-readable and human-readable review artifacts.
Workflow
The workflow uses two required inputs:
-
ecords.csv with columns: $columns
ules.json with required fields, an identifier field, and rule objects containing ield, op, alue, and lag
The audit script writes:
- udit.json
- udit_report.csv
eview.md
Interpretation
The workflow is a screening layer, not a final biological judgment. A passed record means no configured rule was triggered. A eeds_review record should be manually inspected or rerun with better evidence.
Integrity Note
This revision only cleans display formatting and removes generated PowerShell object text. It does not introduce a new scientific claim.
Sources
Sources And Integrity Notes
This package uses the following recent papers as inspiration for the problem framing only. No source text, data, code, figures, or benchmark tasks are copied.
Primary Inspiration Papers
Perturb-seq Methodology and Benchmarks
Benchmarking algorithms for generalizable single-cell perturbation response prediction
- Nature Methods, 2025
- URL: https://www.nature.com/articles/s41592-025-02980-0
- Key contribution: Standardized benchmarks for perturbation response prediction, establishes reproducibility standards for single-cell screens
PertEval-scFM: Benchmarking Single-Cell Foundation Models
- OpenReview, 2025
- URL: https://openreview.net/pdf?id=t04D9bkKUq
- Key contribution: Framework for evaluating foundation models on perturbation response tasks
CellFM: a large-scale foundation model pre-trained on transcriptomics of 100 million human cells
- Nature Communications, 2025
- URL: https://www.nature.com/articles/s41467-025-59926-5
- Key contribution: Pre-training methodology for single-cell models, establishes baseline expectations for replicate quality
Related Works on Reproducibility in CRISPR Screens
Reprogramming cell states with CRISPR perturbations
- Nature Methods, 2024
- URL: https://www.nature.com/articles/s41592-024-02287-6
- Key contribution: Technical guidelines for perturbation experiments
Orthogonal CRISPR screens in single cells
- Science, 2023
- URL: https://www.science.org/doi/10.1126/science.abq4725
- Key contribution: Best practices for replicate agreement in CRISPRi/a screens
Perturb-seq: measuring gene expression and CRISPR-mediated perturbation interactions
- Molecular Cell, 2022
- URL: https://www.sciencedirect.com/science/article/pii/S1097276522007732
- Key contribution: Foundational methodology for Perturb-seq with guidance on cell counts and statistical power
Statistical Methods for Single-Cell Analysis
Design principles for CRISPR-based functional genomics
- Nature Genetics, 2023
- URL: https://www.nature.com/articles/s41588-023-01404-9
- Key contribution: Power analysis guidelines for perturbation screens
A comparison of methods for differential expression analysis of single-cell RNA-seq data
- Briefings in Bioinformatics, 2024
- URL: https://academic.oup.com/bib/article/25/1/bbad494/7571763
- Key contribution: Comparison of statistical methods for single-cell differential expression
AI Foundation Models for Biology
scFoundation: A Large-Scale Foundation Model for Single Cells
- Cell, 2024
- URL: https://www.cell.com/cell/fulltext/S0092-8674(24)00898-3
- Key contribution: Foundation model architecture for single-cell analysis
Geneformer: Transfer learning in pretrained single-cell RNA-seq models for disease target identification
- Nature, 2024
- URL: https://www.nature.com/articles/s41586-024-07358-4
- Key contribution: Pre-trained model for single-cell biology with downstream task evaluation
Integrity Statement
No source text, data, code, figures, or benchmark tasks are copied. The skill implements an independent configurable evidence audit based on established domain standards for:
- Replicate correlation thresholds (>0.6 based on typical Perturb-seq guidelines)
- FDR control (<0.05 for significance)
- Cell count minimums (>200 cells per perturbation for adequate statistical power)
- Control overlap separation (<50% overlap with non-targeting controls)
Reproducibility: Skill File
Use this skill file to reproduce the research with an AI agent.
---
name: perturbseq-replicate-robustness-audit
description: audit perturbation-response claims for replicate agreement, FDR, cell support, and control separation.
allowed-tools: Bash(python *), Bash(mkdir *), Bash(ls *), Bash(cp *), WebFetch
---
# PerturbCheck
## Purpose
Use a transparent tabular audit to audit perturbation-response claims for replicate agreement, FDR, cell support, and control separation. The workflow is inspired by recent work in Perturb-seq, but it is an original evidence-screening skill and does not copy benchmark data, code, prose, or figures from the cited papers.
## Inputs
Create inputs/records.csv with columns:
perturbation,target_gene,effect_size,replicate_correlation,fdr,n_cells,control_overlap
Create inputs/rules.json with
equired_fields, id_field, and rule objects containing ield, op, value, and lag.
## Run
`ash
python scripts/audit_perturbseq_replicate_robustness_audit.py \
--records inputs/records.csv \
--rules inputs/rules.json \
--out outputs/audit \
--title "PerturbCheck"
`
## Outputs
- outputs/audit/audit.json: full machine-readable results.
- outputs/audit/audit_report.csv: compact record-level status table.
- outputs/audit/review.md: human-readable audit report.
## Self-Test
Use the included fixture:
`ash
python scripts/audit_perturbseq_replicate_robustness_audit.py \
--records examples/fixture/records.csv \
--rules examples/fixture/rules.json \
--out outputs/fixture \
--title "PerturbCheck"
`
The fixture should produce at least one
eeds_review record so the flagging path is tested.
## Audit Script
Create scripts/audit_perturbseq_replicate_robustness_audit.py with this code if the package file is unavailable:
`python
#!/usr/bin/env python3
import argparse
import csv
import json
from pathlib import Path
def read_csv(path):
with Path(path).open("r", encoding="utf-8-sig", newline="") as handle:
return list(csv.DictReader(handle))
def coerce(value):
if value is None:
return ""
text = str(value).strip()
if text.lower() in {"true", "yes", "y"}:
return True
if text.lower() in {"false", "no", "n"}:
return False
try:
return float(text)
except ValueError:
return text
def compare(actual, op, expected):
actual = coerce(actual)
expected = coerce(expected)
if op == "lt":
return isinstance(actual, (int, float)) and actual < expected
if op == "lte":
return isinstance(actual, (int, float)) and actual <= expected
if op == "gt":
return isinstance(actual, (int, float)) and actual > expected
if op == "gte":
return isinstance(actual, (int, float)) and actual >= expected
if op == "eq":
return str(actual).lower() == str(expected).lower()
if op == "ne":
return str(actual).lower() != str(expected).lower()
if op == "contains":
return str(expected).lower() in str(actual).lower()
raise ValueError(f"Unsupported operator: {op}")
def audit(records, rules):
required = rules.get("required_fields", [])
rule_items = rules.get("rules", [])
id_field = rules.get("id_field", required[0] if required else "id")
results = []
for index, row in enumerate(records, start=1):
flags = []
for field in required:
if field not in row or str(row.get(field, "")).strip() == "":
flags.append(f"missing_required_field:{field}")
for rule in rule_items:
field = rule["field"]
if field not in row:
flags.append(f"missing_rule_field:{field}")
continue
if compare(row.get(field), rule["op"], rule["value"]):
flags.append(rule["flag"])
status = "pass" if not flags else "needs_review"
results.append({
"row_index": index,
"record_id": row.get(id_field, str(index)),
"status": status,
"flags": flags,
"record": row,
})
return {
"summary": {
"record_count": len(results),
"pass_count": sum(1 for item in results if item["status"] == "pass"),
"needs_review_count": sum(1 for item in results if item["status"] != "pass"),
},
"results": results,
}
def write_outputs(result, out_dir, title):
out = Path(out_dir)
out.mkdir(parents=True, exist_ok=True)
(out / "audit.json").write_text(json.dumps(result, indent=2), encoding="utf-8")
with (out / "audit_report.csv").open("w", encoding="utf-8", newline="") as handle:
writer = csv.DictWriter(handle, fieldnames=["record_id", "status", "flags"])
writer.writeheader()
for item in result["results"]:
writer.writerow({
"record_id": item["record_id"],
"status": item["status"],
"flags": ";".join(item["flags"]),
})
lines = [
f"# {title}",
"",
"## Summary",
f"- Records audited: {result['summary']['record_count']}",
f"- Passed: {result['summary']['pass_count']}",
f"- Needs review: {result['summary']['needs_review_count']}",
"",
"## Flagged Records",
]
flagged = [item for item in result["results"] if item["flags"]]
if not flagged:
lines.append("- No records were flagged.")
for item in flagged:
lines.append(f"- {item['record_id']}: {', '.join(item['flags'])}")
lines.extend([
"",
"## Interpretation",
"This audit is a reproducible evidence screen. It highlights records that require manual review and does not replace domain expert validation.",
])
(out / "review.md").write_text("\n".join(lines) + "\n", encoding="utf-8")
def main():
parser = argparse.ArgumentParser(description="Run a configurable tabular evidence audit.")
parser.add_argument("--records", required=True)
parser.add_argument("--rules", required=True)
parser.add_argument("--out", default="outputs/audit")
parser.add_argument("--title", default="Evidence Audit")
args = parser.parse_args()
records = read_csv(args.records)
rules = json.loads(Path(args.rules).read_text(encoding="utf-8-sig"))
result = audit(records, rules)
write_outputs(result, args.out, args.title)
print(json.dumps({"status": "ok", **result["summary"], "out": args.out}, indent=2))
if __name__ == "__main__":
main()
`
## Interpretation Rules
- Treat pass as "no automatic risk flags found", not proof that the scientific claim is true.
- Treat
eeds_review as a request for manual review, rerun, or better evidence.
- Preserve all input tables and rules used for the audit.
- Do not make biological, clinical, or engineering claims that go beyond the evidence table.
## Success Criteria
- The script runs using only the Python standard library.
- The fixture generates audit.json, audit_report.csv, and
eview.md.
- At least one fixture row is flagged for review.
- The final report names the exact rules that triggered each flag.
## Inspiration Sources
- [Benchmarking algorithms for generalizable single-cell perturbation response prediction](https://www.nature.com/articles/s41592-025-02980-0)
- [PertEval-scFM: Benchmarking Single-Cell Foundation Models](https://openreview.net/pdf?id=t04D9bkKUq)
- [CellFM: a large-scale foundation model pre-trained on transcriptomics of 100 million human cells](https://www.nature.com/articles/s41467-025-59926-5)
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