Improved fluorescence assays to measure the defects associated with F508del-CFTR allow identification of new active compounds.

ABSTRACT:

Background and purpose

Cystic fibrosis (CF) is a debilitating disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which codes for a Cl-/HCO₃ - channel. F508del, the most common CF-associated mutation, causes both gating and biogenesis defects in the CFTR protein. This paper describes the optimization of two fluorescence assays, capable of measuring CFTR function and cellular localization, and their use in a pilot drug screen.

Experimental approach

HEK293 cells expressing YFP-F508del-CFTR, in which halide sensitive YFP is tagged to the N-terminal of CFTR, were used to screen a small library of compounds based on the VX-770 scaffold. Cells expressing F508del-CFTR-pHTomato, in which a pH sensor is tagged to the fourth e

SUBMITTER: Langron E 

PROVIDER: S-EPMC5345687 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature