Isogenic cell models of cystic fibrosis-causing variants in natively expressing pulmonary epithelial cells

Hillary C. Valley, Katherine M. Bukis, Alisa Bell, Yi Cheng, Eric Wong, Nikole J. Jordan, Normand E. Allaire, Andrey Sivachenko, Feng Liang, Hermann Bihler, Philip J Thomas, Jerome Mahiou, Martin Mense

Research output: Contribution to journalArticlepeer-review

59 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)476-483
Number of pages8
JournalJournal of Cystic Fibrosis
Issue number4
StatePublished - Jul 2019


  • CFTR
  • CRISPR/Cas9
  • Cystic Fibrosis
  • Model system
  • Nonsense mutation
  • Premature termination codon

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health
  • Pulmonary and Respiratory Medicine


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