Tracking-seq shows varied unintended effects in CRISPR-Cas9 gene editing

Tracking-seq can identify off-target effects of genome editing tools with low cell input.

• The method is applicable to various genome-editing tools, including Cas9, base editors, and prime editors.
• Tracking-seq detects off-target effects by tracking replication protein A-bound single-stranded DNA.
• The approach is suitable for in vitro, ex vivo, and in vivo applications.
• Heterogeneity in off-target effects is observed between different editor types and cell types.
• Direct measurement in the original system is necessary to assess off-target effects accurately.

Simplified