Title: CRISPR-Cas9 Mediated Gene Editing of the CFTR Locus in Human Bronchial Epithelial Cells
Authors: E. V. Reston, M. T. Williams, S. L. Chen
Affiliations: Department of Genetics, Stanford University, Stanford, CA, USA
Background: Cystic Fibrosis is a monogenic disorder caused by mutations in the CFTR gene. Gene editing technologies like CRISPR-Cas9 offer a potential avenue for permanent correction of these mutations.
Methods: We utilized a lentiviral delivery system to introduce Cas9 and a guide RNA targeting the ΔF508 mutation into primary human bronchial epithelial cells derived from patients with Cystic Fibrosis. Gene correction was assessed via Sanger sequencing and a functional chloride efflux assay.
Results: We achieved a gene correction efficiency of 12±3% in the target cell population. Corrected cells demonstrated a significant restoration of chloride channel function, reaching 35±8% of wild-type levels. Off-target analysis revealed minimal activity at the top three predicted sites.
Conclusion: Our findings demonstrate the feasibility of CRISPR-Cas9 for correcting the primary genetic defect in patient-derived respiratory cells, providing a proof-of-concept for a potential therapeutic strategy for Cystic Fibrosis.
