Production of CFTR-null and CFTR-DeltaF508 heterozygous pigs by adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer.
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.
Rogers
Christopher S
CS
Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, Iowa, USA.
Hao
Yanhong
Y
Rokhlina
Tatiana
T
Samuel
Melissa
M
Stoltz
David A
DA
Li
Yuhong
Y
Petroff
Elena
E
Vermeer
Daniel W
DW
Kabel
Amanda C
AC
Yan
Ziying
Z
Spate
Lee
L
Wax
David
D
Murphy
Clifton N
CN
Rieke
August
A
Whitworth
Kristin
K
Linville
Michael L
ML
Korte
Scott W
SW
Engelhardt
John F
JF
Welsh
Michael J
MJ
Prather
Randall S
RS
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
D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D010649:Phenylalanine