Cell lines expressing cftr and methods of using them
AssigneeChromocell Corporation
InventorsKambiz Shekdar, Jessica Langer, Srinivasan P. Venkatachalan, Dennis J. Sawchuk
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Abstract
Disclosed herein are cells and cell lines that stably express CFTR and methods for using those cells and cell lines. The invention also includes techniques for creating these cells and cell lines. The cells and cell lines of this invention are physiologically relevant. They are highly sensitive and provide consistent and reliable results in cell-based assays.
Claims34
A cell or cell line engineered to stably express cystic fibrosis transmembrane conductance regulator (CFTR).
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A cell engineered to stably express CFTR at a consistent level over time, the cell made by a method comprising the steps of:
- a) providing a plurality of cells that express mRNA(s) encoding the CFTR;
- b) dispersing the cells individually into individual culture vessels, thereby providing a plurality of separate cell cultures;
- c) culturing the cells under a set of desired culture conditions using automated cell culture methods characterized in that the conditions are substantially identical for each of the separate cell cultures, during which culturing the number of cells per separate cell culture is normalized, and wherein the separate cultures are passaged on the same schedule;
- d) assaying the separate cell cultures to measure expression of the CFTR at least twice;
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- e) identifying a separate cell culture that expresses the CFTR at a consistent level in both assays, thereby obtaining said cell.
A method for isolating a cell that endogenously expresses CFTR comprising:
- a) providing a population of cells;
- b) introducing into the cells a molecular beacon that detects expression of CFTR;
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- c) isolating cells that express CFTR.
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A method for increasing the expression level of a CFTR on the plasma membrane of a cell, comprising contacting the cell with a compound of the formula:
- N-{2-[(2-methoxyphenyl)amino]-4′-methyl-4,5′-bi-1,3-thiazol-2′-yl}benzamide hydrobromide.
Dependencies are inferred from the claim text.