Production of CFTR-null and CFTR-ΔF508 heterozygous pigs by adeno-associated virus - Mediated gene targeting and somatic cell nuclear transfer

Christopher S. Rogers, Yanhong Hao, Tatiana Rokhlina, Melissa Samuel, David A. Stoltz, Yuhong Li, Elena Petroff, Daniel W. Vermeer, Amanda C. Kabel, Ziying Yan, Lee Spate, David Wax, Clifton N. Murphy, August Rieke, Kristin Whitworth, Michael L. Linville, Scott W. Korte, John F. Engelhardt, Michael J. Welsh, Randall S. Prather

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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 (ΔF508). 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.

Original languageEnglish
Pages (from-to)1571-1577
Number of pages7
JournalJournal of Clinical Investigation
Volume118
Issue number4
DOIs
StatePublished - 1 Apr 2008

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Dependovirus
Gene Targeting
Cystic Fibrosis
Swine
Inborn Genetic Diseases
Medical Genetics
Genes
Pancreatic Diseases
Mutation
Heterozygote
Biochemistry
Lung Diseases
Oocytes
Anatomy
Animal Models
Fibroblasts
Tissue Donors
Morbidity
Mortality

Cite this

Rogers, Christopher S. ; Hao, Yanhong ; Rokhlina, Tatiana ; Samuel, Melissa ; Stoltz, David A. ; Li, Yuhong ; Petroff, Elena ; Vermeer, Daniel W. ; Kabel, Amanda C. ; Yan, Ziying ; Spate, Lee ; Wax, David ; Murphy, Clifton N. ; Rieke, August ; Whitworth, Kristin ; Linville, Michael L. ; Korte, Scott W. ; Engelhardt, John F. ; Welsh, Michael J. ; Prather, Randall S. / Production of CFTR-null and CFTR-ΔF508 heterozygous pigs by adeno-associated virus - Mediated gene targeting and somatic cell nuclear transfer. In: Journal of Clinical Investigation. 2008 ; Vol. 118, No. 4. pp. 1571-1577.
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title = "Production of CFTR-null and CFTR-ΔF508 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 (ΔF508). 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.",
author = "Rogers, {Christopher S.} and Yanhong Hao and Tatiana Rokhlina and Melissa Samuel and Stoltz, {David A.} and Yuhong Li and Elena Petroff and Vermeer, {Daniel W.} and Kabel, {Amanda C.} and Ziying Yan and Lee Spate and David Wax and Murphy, {Clifton N.} and August Rieke and Kristin Whitworth and Linville, {Michael L.} and Korte, {Scott W.} and Engelhardt, {John F.} and Welsh, {Michael J.} and Prather, {Randall S.}",
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Rogers, CS, Hao, Y, Rokhlina, T, Samuel, M, Stoltz, DA, Li, Y, Petroff, E, Vermeer, DW, Kabel, AC, Yan, Z, Spate, L, Wax, D, Murphy, CN, Rieke, A, Whitworth, K, Linville, ML, Korte, SW, Engelhardt, JF, Welsh, MJ & Prather, RS 2008, 'Production of CFTR-null and CFTR-ΔF508 heterozygous pigs by adeno-associated virus - Mediated gene targeting and somatic cell nuclear transfer', Journal of Clinical Investigation, vol. 118, no. 4, pp. 1571-1577. https://doi.org/10.1172/JCI34773

Production of CFTR-null and CFTR-ΔF508 heterozygous pigs by adeno-associated virus - Mediated gene targeting and somatic cell nuclear transfer. / Rogers, Christopher S.; Hao, Yanhong; Rokhlina, Tatiana; Samuel, Melissa; Stoltz, David A.; Li, Yuhong; Petroff, Elena; Vermeer, Daniel W.; Kabel, Amanda C.; Yan, Ziying; Spate, Lee; Wax, David; Murphy, Clifton N.; Rieke, August; Whitworth, Kristin; Linville, Michael L.; Korte, Scott W.; Engelhardt, John F.; Welsh, Michael J.; Prather, Randall S.

In: Journal of Clinical Investigation, Vol. 118, No. 4, 01.04.2008, p. 1571-1577.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Production of CFTR-null and CFTR-ΔF508 heterozygous pigs by adeno-associated virus - Mediated gene targeting and somatic cell nuclear transfer

AU - Rogers, Christopher S.

AU - Hao, Yanhong

AU - Rokhlina, Tatiana

AU - Samuel, Melissa

AU - Stoltz, David A.

AU - Li, Yuhong

AU - Petroff, Elena

AU - Vermeer, Daniel W.

AU - Kabel, Amanda C.

AU - Yan, Ziying

AU - Spate, Lee

AU - Wax, David

AU - Murphy, Clifton N.

AU - Rieke, August

AU - Whitworth, Kristin

AU - Linville, Michael L.

AU - Korte, Scott W.

AU - Engelhardt, John F.

AU - Welsh, Michael J.

AU - Prather, Randall S.

PY - 2008/4/1

Y1 - 2008/4/1

N2 - 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 (ΔF508). 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.

AB - 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 (ΔF508). 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.

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U2 - 10.1172/JCI34773

DO - 10.1172/JCI34773

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SP - 1571

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SN - 0021-9738

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