CRISPR-Cas9 KO Cell Line Generation And Development Of A Cell-Based Potency Assay For rAAV-FKRP Gene Therapy

A reliable potency assay is essential for demonstrating that rAAV-FKRP gene therapy produces its intended biological effect in LGMDR9. Researchers created CRISPR-Cas9 FKRP knockout muscle cells with complete loss of α-dystroglycan glycosylation, establishing a controlled model for measuring functional restoration. A high-throughput On-Cell Western method was then used to quantify recovery of α-DG glycosylation following treatment with rAAV-FKRP. By calculating the half-maximal effective concentration, or EC50, and comparing results against a reference standard, the assay can support evaluation of product activity, quality, and batch-to-batch consistency.

Explore how this cell-based approach may provide a practical potency assay for rAAV-FKRP products as they advance through development and clinical evaluation.