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Related Concept Videos

Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...

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Related Experiment Video

Updated: Jun 27, 2026

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
09:59

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Published on: March 9, 2015

High-content functional screen to identify proteins that correct F508del-CFTR function.

Agata M Trzcinska-Daneluti1, Diane Ly, Lise Huynh

  • 1Program in Cell Biology, The Hospital for Sick Children, and Biochemistry Department, University of Toronto, Toronto, Ontario M5G 1L7, Canada.

Molecular & Cellular Proteomics : MCP
|December 18, 2008
PubMed
Summary
This summary is machine-generated.

Researchers identified proteins that can correct the F508del-CFTR defect in Cystic Fibrosis. This discovery advances understanding of CFTR protein function and potential therapeutic targets.

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Last Updated: Jun 27, 2026

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
09:59

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

Published on: March 9, 2015

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae
15:12

Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein Expressed in Saccharomyces cerevisiae

Published on: May 10, 2014

Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae
14:56

Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae

Published on: March 10, 2012

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Cystic Fibrosis (CF) is primarily caused by mutations in the CFTR gene.
  • The most common mutation, F508del-CFTR, results in a misfolded protein with a trafficking defect, leading to its degradation.
  • Correcting this defect is crucial for developing effective CF therapies.

Purpose of the Study:

  • To develop and implement a high-content screening assay to identify proteins that can rescue the F508del-CFTR trafficking and function defect.
  • To discover novel protein interactors that modulate CFTR protein processing and function.

Main Methods:

  • Utilized a HEK293 cell line stably expressing F508del-CFTR.
  • Co-expressed approximately 450 unique proteins fused to a YFP reporter.
  • Assessed CFTR function using a chloride efflux assay and high-content imaging.
  • Validated corrector protein function through Western blot and cell surface expression analysis.

Main Results:

  • Identified several known and novel proteins that correct F508del-CFTR function.
  • Validated proteins include STAT1, Endothelin 1, HspA4, and LGALS3/galectin-3.
  • Demonstrated enhanced CFTR maturation and cell surface expression for validated correctors.

Conclusions:

  • High-content screening is effective for identifying proteins that correct CFTR defects.
  • Discovered novel proteins that can modulate F508del-CFTR function, offering potential therapeutic avenues.
  • The developed assay is versatile for various screening approaches, including RNAi and small molecule screens.