An Animal Model for Cystic Fibrosis Made by Gene Targeting

Snouwaert, John N.; Brigman, Kristen; Latour, Anne M.; Malouf, Nadia N.; Boucher, Richard C.; Smithies, Oliver; Koller, Beverly H.

doi:10.1126/science.257.5073.1083

Cited by 860 publications

(625 citation statements)

References 16 publications

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“…For example, the early mouse model of cystic fibrosis never acquired spontaneous and chronic lung infections, but manifested meconium ileus, alteration of mucous and serous glands, and obstruction of gland-like structures. 47 Using the strategy of conditional KO, Hodges et al 48 finally confirmed the role played by cystic fibrosis transmembrane conductance regulator in lung inflammation in mice.…”

Section: Discussionmentioning

confidence: 99%

Subfertility and growth restriction in a new galactose-1 phosphate uridylyltransferase (GALT) - deficient mouse model

et al. 2014

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The first GalT gene knockout (KO) mouse model for Classic Galactosemia (OMIM 230400) accumulated some galactose and its metabolites upon galactose challenge, but was seemingly fertile and symptom free. Here we constructed a new GalT genetrapped mouse model by injecting GalT gene-trapped mouse embryonic stem cells into blastocysts, which were later implanted into pseudo-pregnant females. High percentage GalT gene-trapped chimera obtained were used to generate heterozygous and subsequently, homozygous GalT gene-trapped mice. Biochemical assays confirmed total absence of galactose-1 phosphate uridylyltransferase (GALT) activity in the homozygotes. Although the homozygous GalT gene-trapped females could conceive and give birth when fed with normal chow, they had smaller litter size (P ¼ 0.02) and longer time-to-pregnancy (P ¼ 0.013) than their wild-type littermates. Follicle-stimulating hormone levels of the mutant female mice were not significantly different from the age-matched, wild-type females, but histological examination of the ovaries revealed fewer follicles in the homozygous mutants (P ¼ 0.007). Administration of a high-galactose (40% w/w) diet to lactating homozygous GalT gene-trapped females led to lethality in over 70% of the homozygous GalT gene-trapped pups before weaning. Cerebral edema, abnormal changes in the Purkinje and the outer granular cell layers of the cerebellum, as well as lower blood GSH/GSSG ratio were identified in the galactose-intoxicated pups. Finally, reduced growth was observed in GalT gene-trapped pups fed with normal chow and all pups fed with high-galactose (20% w/w) diet. This new mouse model presents several of the complications of Classic Galactosemia and will be useful to investigate pathogenesis and new therapies.

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Section: Discussionmentioning

confidence: 99%

Subfertility and growth restriction in a new galactose-1 phosphate uridylyltransferase (GALT) - deficient mouse model

et al. 2014

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“…Mice homozygous for the S489X (B6.129P2-Cftr tm1Unc ) mutation of the CFTR gene, congenic (n ϭ 10 generations) onto C57BL/6J background, (18) and their normal littermates, designated as CFTR Ϫ/Ϫ and CFTR ϩ/ϩ , respectively, were bred in our Animal Core facility. The genotype of individual animals was established by PCR amplification of tail snip genomic DNA (19,20).…”

Section: Micementioning

confidence: 99%

Functional IL-10 Deficiency in the Lung of Cystic Fibrosis (cftr−/−) and IL-10 Knockout Mice Causes Increased Expression and Function of B7 Costimulatory Molecules on Alveolar Macrophages

Šoltýs

Bonfield

Chmiel

et al. 2002

The Journal of Immunology

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Alveolar macrophages are poor APCs that only minimally express B7 costimulatory molecules. Because our previous data suggest that bronchial epithelial cells constitutively secrete IL-10, and IL-10 inhibits B7 expression in vitro, we hypothesized that this IL-10 is responsible for suppressing B7 expression on macrophages that enter the airways. Furthermore, because we have shown that cystic fibrosis (CF) lungs are deficient in IL-10, we hypothesized that bronchoalveolar macrophages (BALMs) from cystic fibrosis transmembrane conductance regulator (CFTR)−/− as well as IL-10−/− mice might express increased B7. Immunofluorescence for B7 was positive on BALMs from CF patients and CFTR−/− and IL-10−/− mice, but was negative on controls. FACS showed that 63.9% of BALMs from IL-10−/− mice were B7-1 positive, as were 67.4% of BALMs from CFTR−/− mice, whereas <7% of BALMs from wild-type controls were positive. Using BALMs to costimulate splenic T cells with anti-CD3 as a mitogen showed 9202 ± 2107 cpm [3H]thymidine incorporation for BALMs from IL-10−/− mice and 4082 ± 1036 cpm for BALMs from CFTR−/− mice, but <200 cpm with BALMs from either type of +/+ mouse. Treatment of CFTR−/− mice with recombinant mouse IL-10 reduced the B7 expression and costimulatory activity of the BALMs. These data suggest that the IL-10 secreted in the healthy lung may be responsible for the absence of B7 and poor costimulatory activity of BALMs and that reductions of pulmonary IL-10 in CF may enhance B7 expression and local immune responses.

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“…Animal models of cystic fibrosis have now been developed in mice [26][27][28] and therefore it is important to establish the effect of pulmonary surfactant on in vivo plasmid DNA transfection in the mouse respiratory epithelium. We have evaluated the transfection of two different DNA expression plasmids coding for chloram-phenicol acetyltransferase (CAT) and luciferase (Luc), in vivo in mice after nasal administration.…”

Section: Introductionmentioning

confidence: 99%

The effect of synthetic surfactant Exosurf on gene transfer in mouse lung in vivo

et al. 1998

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Gene transfer in the lung holds promise for the treatment for 5 days thereafter. When DNA was delivered in a 50% of diseases such as pulmonary fibrosis, cystic fibrosis and suspension of Exosurf, the expression of either CAT or asthma. Pulmonary surfactant has been reported to Luc was significantly reduced by 89.6 ± 1.4% and enhance expression from endobronchial, adenovirus-82.7 ± 10.5%, respectively. The decrease in Luc mediated gene transfer in experimental animals. This study expression was closely correlated (r = 0.99, P Ͻ 0.001) to examines the effect of exogenous synthetic surfactant log concentration of surfactant in the plasmid buffer sol-(Exosurf) on gene expression from naked plasmid DNA ution (IC 50 = 8.6%). CAT expression was not altered when administered endobronchially to adult mice. Transfection surfactant was administered either 2 h before or after plasefficiency was evaluated by quantifying the expression of mid DNA instillation. Examination of the components of chloramphenicol acetyltransferase (CAT) and luciferase Exosurf revealed that two compounds, DPPC and tylox-(Luc) genes in the lung. Endobronchial administration of apol, showed inhibitory effects on CAT expression. Howeither CAT or Luc expression plasmid DNA resulted in ever, the inhibition caused by Exosurf appeared greater detectable concentrations of each reporter protein. CAT than that of either component. Our results suggest that the expression from plasmid DNA was monitored after endolung surfactant is a barrier to transfection of the endobronchial administration with the maximal expression bronchial airway and may be partly responsible for the low observed at 3-5 days after administration and decreasing expression of exogenous DNA in vivo in the bronchial tree.

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An Animal Model for Cystic Fibrosis Made by Gene Targeting

Cited by 860 publications

References 16 publications

Subfertility and growth restriction in a new galactose-1 phosphate uridylyltransferase (GALT) - deficient mouse model

Subfertility and growth restriction in a new galactose-1 phosphate uridylyltransferase (GALT) - deficient mouse model

Functional IL-10 Deficiency in the Lung of Cystic Fibrosis (cftr−/−) and IL-10 Knockout Mice Causes Increased Expression and Function of B7 Costimulatory Molecules on Alveolar Macrophages

The effect of synthetic surfactant Exosurf on gene transfer in mouse lung in vivo

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