CRISPRa Guide RNA Cloning Protocol for High-Throughput Screening in C. elegans

A 36.7 KB Excel file by Shusen Zhu, last updated May 20, 2026, describes a streamlined cloning protocol for CRISPR activation. The protocol details a pooled, one-step dual gRNA cloning method designed to accelerate the construction of gRNA expression vectors. It was tested with 126 gRNA inserts, achieving 42%–56% coverage per cloning round, and is intended for large-scale gene activation studies in C. elegans.

Use Cases

• Designing high-throughput CRISPRa screens based on the described pooled cloning workflow.
• Optimizing laboratory protocols for gRNA vector construction based on the reported efficiency metrics.
• Conducting phenotypic or lifespan analysis in C. elegans using the generated gRNA libraries.

Strengths

• Protocol tested with a specific batch of 126 gRNA inserts.
• Reports cloning coverage of 42%–56% per round in a test case.
• Released under a permissive CC-BY-4.0 license.

Limitations

• Row count and specific column-level data are unknown, limiting suitability assessment.
• Description metadata is limited; actual data quality requires manual inspection after download.
• The 36.7 KB file size suggests a very limited scope, likely containing protocol details rather than large-scale experimental results.

Provenance

Source

figshare, author Shusen Zhu.

Collection Method

Likely contains protocol details and results from a laboratory method development study.

Freshness

Last updated 2026-05-20 05:56:00; freshness should be verified.