[HTML][HTML] Creation and characterization of an airway epithelial cell line for stable expression of CFTR variants
LB Gottschalk, B Vecchio-Pagan, N Sharma… - Journal of Cystic …, 2016 - Elsevier
Journal of Cystic Fibrosis, 2016•Elsevier
Background Analysis of the functional consequences and treatment response of rare CFTR
variants is challenging due to the limited availability of primary airways cells. Methods 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 …
variants is challenging due to the limited availability of primary airways cells. Methods 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 …
Background
Analysis of the functional consequences and treatment response of rare CFTR variants is challenging due to the limited availability of primary airways cells.
Methods
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.
Results
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.
Conclusion
CF8Flp cells provide a viable substitute for primary CF airway cells for the analysis of CFTR variants in a native context.
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