Zoom Link
https://muw.zoom.us/j/96073587295
Department
Sciences & Mathematics
Format of Presentation
Oral Presentation
Description
CFTR (Cystic Fibrosis Transmembrane-conductance Regulator) is a plasma membrane protein that functions as a chloride ion channel on many epithelial cells. There are over 1000 mutations that affect the function of this protein; however, the most common mutation is DF508. CFTR mutations target many secretory organs like pancreas and lungs causing the genetic disease cystic fibrosis (CF), common among Caucasians of northern European origin. Detection of endogenous CFTR expression is possible by a difficult and costly immunoprecipitation method but not by the most used western blotting method. Therefore, the goal of this project was to detect the endogenous CFTR expression in pancreatic cell lines CFPAC (expressing DF508-CFTR) and Capan-1 (expressing wildtype-CFTR) by the western blotting. With the help of a newly developed western blotting method [Heda et al, BioTechniques, 68(6), 319-325, 2020] and modern sensitive detection tools, I am able to detect the endogenous expression of DF508 and wild-type CFTR. Endogenous CFTR detection by this improved western blotting method, however, required a large amount of protein samples. Specificity of the CFTR expression was confirmed by the peptide inhibition experiment. This new detection method may serve as a diagnostic tool for the detection of CFTR in CF specimen.
Recommended Citation
Amezquita, Zithlaly, "Endogenous CFTR Expression in Human Epithelial Cell Lines" (2022). Undergraduate Research Conference. 1.
https://athenacommons.muw.edu/urc/2022/STEM/1
Included in
Biochemistry Commons, Biology Commons, Molecular Biology Commons
Endogenous CFTR Expression in Human Epithelial Cell Lines
CFTR (Cystic Fibrosis Transmembrane-conductance Regulator) is a plasma membrane protein that functions as a chloride ion channel on many epithelial cells. There are over 1000 mutations that affect the function of this protein; however, the most common mutation is DF508. CFTR mutations target many secretory organs like pancreas and lungs causing the genetic disease cystic fibrosis (CF), common among Caucasians of northern European origin. Detection of endogenous CFTR expression is possible by a difficult and costly immunoprecipitation method but not by the most used western blotting method. Therefore, the goal of this project was to detect the endogenous CFTR expression in pancreatic cell lines CFPAC (expressing DF508-CFTR) and Capan-1 (expressing wildtype-CFTR) by the western blotting. With the help of a newly developed western blotting method [Heda et al, BioTechniques, 68(6), 319-325, 2020] and modern sensitive detection tools, I am able to detect the endogenous expression of DF508 and wild-type CFTR. Endogenous CFTR detection by this improved western blotting method, however, required a large amount of protein samples. Specificity of the CFTR expression was confirmed by the peptide inhibition experiment. This new detection method may serve as a diagnostic tool for the detection of CFTR in CF specimen.
https://athenacommons.muw.edu/urc/2022/STEM/1