AviTag-seq unifies nucleotide-resolution maps of CRISPR off-targets and AAV vector integrations

Description

Jia-Xin Li and eleven colleagues introduced in 2026 a single sequencing method that maps two safety risks of gene-editing therapies. Regulators need to know both where an editor cuts the wrong DNA sites and where a delivery virus inserts itself, yet current break-dependent assays miss editors that nick rather than cut. Tag polarity limits capture. AviTag-seq repurposes the hairpin ends of adeno-associated virus vectors, called inverted terminal repeats, as tags.

Because the hairpin structure sidesteps polarity problems, one primer pair gives high-sensitivity detection. The method catches off-target events from both editor types that assays overlook. In mouse liver it outperformed a method at profiling Pcsk9 off-targets while mapping virus integration sites. Unlike laboratory results, virus vectors in living liver preferred active gene promoters, a risk for liver-directed therapies. The authors offer the platform as a regulatory-grade safety check.

Catalog record

Source: https://europepmc.org/articles/PMC13473653