A powerful method for in vitro selection of normal versus cystic fibrosis airway epithelial cells.
In preparation for a gene therapy approach to cystic fibrosis involving the precise repair of mutations on the CF gene by homologous recombination, we developed a method that would allow for selection of the CFTR+ cells originated in gene targeting experiments on CFTR- cells in vitro. The method is based on the differential sensitivity we observed between CFTR+ and CFTR- cells to agents stimulating cyclic adenosine monophosphate (cAMP). Controlled treatment with epinephrine or forskolin allows for selectively killing CFTR- cells. The efficiency of the selection method would make it suitable for rescuing the few corrected cells originated from rare homologous recombination events.
Vega
M A
MA
Laboratoire de Genetique Moleculaire, U.91- INSERM, Hopital H. Mondor, Creteil (Paris), France.
Goossens
M
M
Besmond
C
C
D002461:Cell Line, Transformed; D002469:Cell Separation / Q000379:methods*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004837:Epinephrine / Q000494:pharmacology; D004848:Epithelium / Q000473:pathology; D018390:Gene Targeting; D015316:Genetic Therapy; D006801:Humans; D066298:In Vitro Techniques; D012137:Respiratory System / Q000473:pathology*
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; D000242:Cyclic AMP; D004837:Epinephrine