title: Long-term survival of the exon 10 insertional cystic fibrosis mutant mouse is a consequence of low level residual wild-type Cftr gene expression.
abstract: Recently we have created a mouse model of cystic fibrosis (CF) by insertional gene targeting to exon 10. In common with CF subjects, this model displays a low incidence of meconium ileus. This contrasts strikingly with the very high level of fatal intestinal obstruction in the three other CF mouse models so far described. We investigate here the molecular basis of this difference in phenotype. We show that the partial duplication consequent upon insertional gene targeting allows exon skipping and aberrant splicing to produce normal Cftr mRNA, but at levels greatly reduced compared with wild-type mice. Furthermore, instead of the predicted mutant Cftr transcript, a novel mRNA is produced that utilizes cryptic splice sites in the disrupting plasmid sequence. However, we have previously shown that these mice display the ion transport defect characteristic of CF, and mutant animals can be distinguished from their normal littermates on this basis. Consistent with this, residual CFTR function has recently been observed for several "mild" mutations in CF individuals who display pancreatic sufficiency but still develop lung disease. We conclude that (i) residual wild-type mRNA in the exon 10 insertional mutant mouse ameliorates the severity of the intestinal phenotype observed in the absolute "null" CF mice, (ii) the presence of low-level residual wild-type Cftr mRNA does not correct the CF ion transport defect, and (iii) the long-term survival of this insertional mutant mouse provides the opportunity to address the factors important in development of lung disease.
authors.affiliation: Medical Research Council Human Genetics, Unit, Western General Hospital, Edinburgh, UK.
authors.forename: J R
authors.initials: JR
authors.lastname: Dorin
authors.forename: B J
authors.initials: BJ
authors.lastname: Stevenson
authors.forename: S
authors.initials: S
authors.lastname: Fleming
authors.forename: E W
authors.initials: EW
authors.lastname: Alton
authors.forename: P
authors.initials: P
authors.lastname: Dickinson
authors.forename: D J
authors.initials: DJ
authors.lastname: Porteous
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018076:DNA, Complementary / Q000235:genetics; D004195:Disease Models, Animal; D005091:Exons; D015870:Gene Expression; D007415:Intestinal Obstruction / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008168:Lung / Q000378:metabolism; D008470:Meconium; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger