[HTML][HTML] Isogenic cell models of cystic fibrosis-causing variants in natively expressing pulmonary epithelial cells
HC Valley, KM Bukis, A Bell, Y Cheng, E Wong… - Journal of Cystic …, 2019 - Elsevier
HC Valley, KM Bukis, A Bell, Y Cheng, E Wong, NJ Jordan, NE Allaire, A Sivachenko…
Journal of Cystic Fibrosis, 2019•ElsevierBackground Assessment of approved drugs and developmental drug candidates for rare
cystic fibrosis (CF)-causing variants of the Cystic Fibrosis Transmembrane Conductance
Regulator (CFTR) requires abundant material from relevant models. Methods Isogenic cell
lines harboring CFTR variants in the native genomic context were created through the
development and utilization of a footprint-less, CRISPR/Cas9 gene editing pipeline in
16HBE14o-immortalized bronchial epithelial cells. Results Isogenic, homozygous cell lines …
cystic fibrosis (CF)-causing variants of the Cystic Fibrosis Transmembrane Conductance
Regulator (CFTR) requires abundant material from relevant models. Methods Isogenic cell
lines harboring CFTR variants in the native genomic context were created through the
development and utilization of a footprint-less, CRISPR/Cas9 gene editing pipeline in
16HBE14o-immortalized bronchial epithelial cells. Results Isogenic, homozygous cell lines …
Background
Assessment of approved drugs and developmental drug candidates for rare cystic fibrosis (CF)-causing variants of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) requires abundant material from relevant models.
Methods
Isogenic cell lines harboring CFTR variants in the native genomic context were created through the development and utilization of a footprint-less, CRISPR/Cas9 gene editing pipeline in 16HBE14o- immortalized bronchial epithelial cells.
Results
Isogenic, homozygous cell lines for three CFTR variants (F508del and the two most common CF-causing nonsense variants, G542X and W1282X) were established and characterized. The F508del model recapitulates the known molecular pathology and pharmacology. The two models of nonsense variants (G542X and W1282X) are sensitive to Nonsense Mediated mRNA Decay (NMD) and responsive to reference compounds that inhibit NMD and promote ribosomal readthrough.
Conclusions
We present a versatile, efficient gene editing pipeline that can be used to create CFTR variants in the native genomic context and the utilization of this pipeline to create homozygous cell models for the CF-causing variants F508del, G542X, and W1282X. The resulting cell lines provide a virtually unlimited source of material with specific pathogenic mutations that can be used in a variety of assays, including functional assays.
Elsevier