Transgenic Rabbits as a Model Organism for Production of Human Clotting Factor VIII

  • Dusan Vasicek
  • Peter Chrenek
  • Alexander Makarevich
  • Miroslav Bauer
  • Rastislav Jurcik
  • Karin Suvegova
  • Jan Rafay
  • Jozef Bulla
  • Ladislav Hetenyi
  • Jeff Erickson
  • Rekha K. Paleyanda
Conference paper

Abstract

Human clotting factor VIII (hFVIII) is a very complex and large protein whose expression is difficult, as hFVIII requires extensive post-translational modification to be biologically active. This paper reports the generation of transgenic rabbits as a model species for testing the expression of hFVIII in the mammary gland. For micro-injection, a fusion gene construct was used, consisting of 2.5 kb murine whey acidic protein (mWAP) promoter, 7.2 kb cDNA of hFVIII, and 4.6 kb of 3′ flanking sequences of the mWAP gene. from 130 micro-injected zygotes transferred into recipients, 30 offspring were delivered. The pups were screened for the transgene by PCR, using DNA isolated from the ear, and results were confirmed by Southern blot analysis. The transgene was identified in one female founder animal, and it was transmitted to the offspring in a Mendelian fashion, thus demonstrating stable integration of the gene construct into the germline of the transgenic rabbits.

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References

  1. Chrenek, P., Makarevich, A., Vasicek, D., Laurincik, J., Bulla, J., Gajarska, T. & Rafay, J. 1998. Effects of superovulation, culture and micro-injection on development of rabbit embryos in vitro. Theriogenology, 50: 659–666.CrossRefPubMedGoogle Scholar
  2. Dove, A. 2000. Milking the genome for profit. Nature Biotechnology, 18: 1045–1048.PubMedGoogle Scholar
  3. Hiripi, L., Makovics, F., Halter, R., Baranyi, M., Paul, D., Carnwath, J.W., Bösze, Z.S. & Niemann, H. 2003. Expression of active human blood clotting factor VIII in the mammary gland of transgenic rabbits. DNA and Cell Biology, 22: 41–45.CrossRefPubMedGoogle Scholar
  4. Jennes, R. 1974. The composition of milk. pp. 3–105, in: Lactation, A Comprehensive Treatise. New York, NY: Academic Press.Google Scholar
  5. Niemann, H., Halter, R., Carnwath, J.W., Herrmann, D., Lemme, E. & Paul, D. 1999. Expression of human blood clotting factor VIII in the mammary gland of transgenic sheep. Transgenic Research, 8: 137–149.CrossRefGoogle Scholar
  6. Paleyanda, R.K., Velander, W.H., Lee, T.K., Scandella, D.H., Gwazdauskas, F.G., Knight, J.W., Hoiyer, L.W., Drohan, W.N. & Lubon, H. 1997. Transgenic pigs produce functional human factor VIII in milk. Nature Biotechnology, 15: 971–975.CrossRefPubMedGoogle Scholar

Copyright information

© IAEA 2005

Authors and Affiliations

  • Dusan Vasicek
    • 1
  • Peter Chrenek
    • 1
  • Alexander Makarevich
    • 1
  • Miroslav Bauer
    • 1
  • Rastislav Jurcik
    • 1
  • Karin Suvegova
    • 1
  • Jan Rafay
    • 1
  • Jozef Bulla
    • 1
  • Ladislav Hetenyi
    • 1
  • Jeff Erickson
    • 2
  • Rekha K. Paleyanda
    • 2
  1. 1.Research Institute of Animal ProductionNitraSlovak Republic
  2. 2.American Integrated Biologics, Inc.E. WoodstockUSA

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