title: Production of CFTR-null and CFTR-DeltaF508 heterozygous pigs by adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer.
abstract: Progress toward understanding the pathogenesis of cystic fibrosis (CF) and developing effective therapies has been hampered by lack of a relevant animal model. CF mice fail to develop the lung and pancreatic disease that cause most of the morbidity and mortality in patients with CF. Pigs may be better animals than mice in which to model human genetic diseases because their anatomy, biochemistry, physiology, size, and genetics are more similar to those of humans. However, to date, gene-targeted mammalian models of human genetic disease have not been reported for any species other than mice. Here we describe the first steps toward the generation of a pig model of CF. We used recombinant adeno-associated virus (rAAV) vectors to deliver genetic constructs targeting the CF transmembrane conductance receptor (CFTR) gene to pig fetal fibroblasts. We generated cells with the CFTR gene either disrupted or containing the most common CF-associated mutation (DeltaF508). These cells were used as nuclear donors for somatic cell nuclear transfer to porcine oocytes. We thereby generated heterozygote male piglets with each mutation. These pigs should be of value in producing new models of CF. In addition, because gene-modified mice often fail to replicate human diseases, this approach could be used to generate models of other human genetic diseases in species other than mice.
authors.affiliation: Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, Iowa, USA.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Rogers
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Hao
authors.forename: Tatiana
authors.initials: T
authors.lastname: Rokhlina
authors.forename: Melissa
authors.initials: M
authors.lastname: Samuel
authors.forename: David A
authors.initials: DA
authors.lastname: Stoltz
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Li
authors.forename: Elena
authors.initials: E
authors.lastname: Petroff
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Vermeer
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Kabel
authors.forename: Ziying
authors.initials: Z
authors.lastname: Yan
authors.forename: Lee
authors.initials: L
authors.lastname: Spate
authors.forename: David
authors.initials: D
authors.lastname: Wax
authors.forename: Clifton N
authors.initials: CN
authors.lastname: Murphy
authors.forename: August
authors.initials: A
authors.lastname: Rieke
authors.forename: Kristin
authors.initials: K
authors.lastname: Whitworth
authors.forename: Michael L
authors.initials: ML
authors.lastname: Linville
authors.forename: Scott W
authors.initials: SW
authors.lastname: Korte
authors.forename: John F
authors.initials: JF
authors.lastname: Engelhardt
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Welsh
authors.forename: Randall S
authors.initials: RS
authors.lastname: Prather
mesh_terms: D000483:Alleles; D000818:Animals; D030801:Animals, Genetically Modified; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics*; D005347:Fibroblasts; D005786:Gene Expression Regulation; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D016678:Genome / Q000235:genetics; D006579:Heterozygote; D009154:Mutation / Q000235:genetics; D053652:Nuclear Transfer Techniques; D010649:Phenylalanine / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D013552:Swine
chemical_list: D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D010649:Phenylalanine