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Sequential targeting of CFTR by BAC vectors generates a novel pig model of cystic fibrosis

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Abstract

Cystic fibrosis (CF) is the most common lethal inherited disease in Caucasians and is caused by mutations in the CFTR gene. The disease is incurable and medical treatment is limited to the amelioration of symptoms or secondary complications. A comprehensive understanding of the disease mechanisms and the development of novel treatment options require appropriate animal models. Existing CF mouse models fail to reflect important aspects of human CF. We thus generated a CF pig model by inactivating the CFTR gene in primary porcine cells by sequential targeting using modified bacterial artificial chromosome vectors. These cells were then used to generate homozygous CFTR mutant piglets by somatic cell nuclear transfer. The homozygous CFTR mutants lack CFTR protein expression and display severe malformations in the intestine, respiratory tract, pancreas, liver, gallbladder, and male reproductive tract. These phenotypic abnormalities closely resemble both the human CF pathology as well as alterations observed in a recently published CF pig model which was generated by a different gene targeting strategy. Our new CF pig model underlines the value of the CFTR-deficient pig for gaining new insight into the disease mechanisms of CF and for the development and evaluation of new therapeutic strategies. This model will furthermore increase the availability of CF pigs to the scientific community.

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Acknowledgments

This study was supported by the Mukoviszidose Institut gemeinnützige Gesellschaft für Forschung und Therapieentwicklung mbH and - in part - by the Bundesministerium für Bildung und Forschung (Leading Edge Cluster Munich: m4, Personalized Medicine and Targeted Therapies). We thank Tuna Güngör, Christian Erdle, Siegfried Elsner, Jana Enders, Gabriele Hahn, Alexandra Harder, and Petra Nehrig for excellent technical support, and Lynda Ostedgaard and Michael J. Welsh for donation of the pCFTR clone.

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Authors and Affiliations

  1. Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Straße 25, 81377, Munich, Germany

    N. Klymiuk, K. Kraehe, A. Wuensch, C. Kröner, A. Richter, B. Kessler, M. Kurome & E. Wolf

  2. Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163, Berlin, Germany

    L. Mundhenk, S. Plog & A. D. Gruber

  3. Institute of Veterinary Pathology, Ludwig-Maximilians-Universität München, Veterinärstraße 13, 80539, Munich, Germany

    D. Emrich & M. C. Langenmayer

  4. Department of Pediatric Surgery, Dr. von Hauner Children’s Hospital, Ludwig-Maximilians-Universität München, Lindwurmstraße 4, 80337, Munich, Germany

    M. Stehr

  5. Division of Neonatology, Dr. von Hauner Children’s Hospital, Ludwig-Maximilians-Universität München, Lindwurmstraße 4, 80337, Munich, Germany

    A. Holzinger

  6. Clinic for Swine, Ludwig-Maximilians-Universität München, Sonnenstraße 16, 85764, Oberschleißheim, Germany

    M. Eddicks & K. Heinritzi

  7. Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki, 214-8571, Japan

    H. Nagashima

Authors
  1. N. Klymiuk
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  2. L. Mundhenk
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  3. K. Kraehe
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  4. A. Wuensch
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  5. S. Plog
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  6. D. Emrich
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  7. M. C. Langenmayer
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  8. M. Stehr
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  9. A. Holzinger
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  10. C. Kröner
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  11. A. Richter
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  12. B. Kessler
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  13. M. Kurome
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  14. M. Eddicks
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  15. H. Nagashima
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  16. K. Heinritzi
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  17. A. D. Gruber
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  18. E. Wolf
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Corresponding authors

Correspondence to N. Klymiuk or E. Wolf.

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Klymiuk, N., Mundhenk, L., Kraehe, K. et al. Sequential targeting of CFTR by BAC vectors generates a novel pig model of cystic fibrosis. J Mol Med 90, 597–608 (2012). https://doi.org/10.1007/s00109-011-0839-y

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  • DOI: https://doi.org/10.1007/s00109-011-0839-y

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