CFTR regions containing duodenum specific DNase I hypersensitive sites drive expression in intestinal crypt cells but not in fibroblasts.
We have investigated CFTR specific intestinal expression by transfection assays in mouse cultured fibroblasts and transimmortalized intestinal crypt m-ICc12 cells using the beta-galactosidase gene linked to rat CFTR non-coding regions. Two constructs were studied, one encompassing a 5.3 kb region 5' to the gene where numerous duodenum-specific DNase I hypersensitive sites (DHSs) were previously mapped and the other including a 1.3 kb 3' region in which novel DHSs had been identified. In transient transfection assays, transgenes were expressed in m-ICc12 cells but not in fibroblasts. In m-ICc12 cells, the pattern of expression of the chromosomally integrated transgenes paralleled the endogenous expression of CFTR and beta-galactosidase activity was detected in cells containing villin and forming domes. Thus, a 6.6 kb region encompassing 5' and 3' non-coding parts of rat CFTR is able to drive specific expression of a reporter gene in cultured mouse intestinal cells having kept a crypt phenotype.
Dixméras
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I
Institut National de la Santé et de la Recherche Médicale (INSERM) unité 458, Hôpital Robert Debré, Paris, France.
Lapouméroulie
C
C
Tallec
L P
LP
Bens
M
M
Elion
J
J
Vandewalle
A
A
Denamur
E
E
D016475:3T3 Cells; D000818:Animals; D003550:Cystic Fibrosis / Q000235:genetics / Q000503:physiopathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D003850:Deoxyribonuclease I / Q000235:genetics*; D005786:Gene Expression Regulation; D007422:Intestines / Q000502:physiology*; D051379:Mice; D051381:Rats; D014162:Transfection
D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D003850:Deoxyribonuclease I