Phenotyping of Rare CFTR Mutations Reveals Distinct Trafficking and Functional Defects.

ABSTRACT: Background. The most common CFTR mutation, F508del, presents with multiple cellular defects. However, the possible multiple defects caused by many rarer CFTR mutations are not well studied. We investigated four rare CFTR mutations E60K, G85E, E92K and A455E against well-characterized mutations, F508del and G551D, and their responses to corrector VX-809 and/or potentiator VX-770. Methods. Using complementary assays in HEK293T stable cell lines, we determined maturation by Western blotting, trafficking by flow cytometry using extracellular 3HA-tagged CFTR, and function by halide-sensitive YFP quenching. In the forskolin-induced swelling assay in intestinal organoids, we validated the effect of tagged versus endogenous CFTR. Results. Treatment with VX

SUBMITTER: Ensinck M 

PROVIDER: S-EPMC7140603 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature