title: Creation and characterization of an airway epithelial cell line for stable expression of CFTR variants.
abstract: Analysis of the functional consequences and treatment response of rare CFTR variants is challenging due to the limited availability of primary airways cells.
  
  
  
  A Flp recombination target (FRT) site for stable expression of CFTR was incorporated into an immortalized CF bronchial epithelial cell line (CFBE41o-). CFTR cDNA was integrated into the FRT site. Expression was evaluated by western blotting and confocal microscopy and function measured by short circuit current. RNA sequencing was used to compare the transcriptional profile of the resulting CF8Flp cell line to primary cells and tissues.
  
  
  
  Functional CFTR was expressed from integrated cDNA at the FRT site of the CF8Flp cell line at levels comparable to that seen in native airway cells. CF8Flp cells expressing WT-CFTR have a stable transcriptome comparable to that of primary cultured airway epithelial cells, including genes that play key roles in CFTR pathways.
  
  
  
  CF8Flp cells provide a viable substitute for primary CF airway cells for the analysis of CFTR variants in a native context.
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Laura B
authors.initials: LB
authors.lastname: Gottschalk
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Briana
authors.initials: B
authors.lastname: Vecchio-Pagan
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Sharma
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Sangwoo T
authors.initials: ST
authors.lastname: Han
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Arianna
authors.initials: A
authors.lastname: Franca
authors.affiliation: Cytogenetics Laboratory, Kennedy Krieger Institute, Baltimore, MD, United States.
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Wohler
authors.affiliation: Cytogenetics Laboratory, Kennedy Krieger Institute, Baltimore, MD, United States.
authors.forename: Denise A S
authors.initials: DA
authors.lastname: Batista
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States; Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
authors.forename: Loyal A
authors.initials: LA
authors.lastname: Goff
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States; Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD, United States. Electronic address: gcutting@jhmi.edu.
authors.forename: Garry R
authors.initials: GR
authors.lastname: Cutting
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D006801:Humans; D059645:Mutation Rate; D020545:Respiratory Mucosa / Q000473:pathology*
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFBE; CFTR; Cystic fibrosis model; Ivacaftor; RNA sequencing