A Versatile Reporter Platform for Evaluating HDR- and NHEJ-Based Genome Editing in Airway Epithelial Cell Cultures Using an rAAV Vector.

ABSTRACT: Therapeutic gene editing strategies utilize endogenous DNA repair pathways-nonhomologous end joining (NHEJ) or homology-directed repair (HDR)-to introduce targeted genomic modifications. Because HDR is restricted to dividing cells, whereas NHEJ functions in both dividing and non-dividing cells, NHEJ-based approaches are better suited for in vivo gene editing in the largely post-mitotic airway epithelium. Homology-independent targeted insertion (HITI), an NHEJ-based method, offers a promising strategy for cystic fibrosis (CF) gene therapy. Here, we applied HITI to drive the expression of a promoterless reporter through an exon trap strategy in both proliferating airway basal cells and well-differentiated primary airway epithelial cultures derived from transgenic ROSA^mTmG ferrets.

SUBMITTER: Park SY 

PROVIDER: S-EPMC12197413 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature