Transgenic Research

, Volume 21, Issue 3, pp 605–618

The creation of transgenic pigs expressing human proteins using BAC-derived, full-length genes and intracytoplasmic sperm injection-mediated gene transfer

  • Masahito Watanabe
  • Mayuko Kurome
  • Hitomi Matsunari
  • Kazuaki Nakano
  • Kazuhiro Umeyema
  • Akira Shiota
  • Hiromitsu Nakauchi
  • Hiroshi Nagashima
Original Paper

DOI: 10.1007/s11248-011-9561-3

Cite this article as:
Watanabe, M., Kurome, M., Matsunari, H. et al. Transgenic Res (2012) 21: 605. doi:10.1007/s11248-011-9561-3

Abstract

In most transgenic (Tg) animals created to date, a transgene consisting of the minimum promoter region linked to a cDNA has been used. However, transgenes on small plasmids are susceptible to the position effect, increasing the difficulty of controlling transgene expression. In this study, we attempted to create Tg pigs by intracytoplasmic sperm injection-mediated gene transfer (ICSI-MGT) using two large genomic transgenes derived from a bacterial artificial chromosome (BAC) containing the full genomic region encoding two human proteins, type I collagen and albumin. The production efficiencies (Tg piglets/live offspring) of type I collagen and albumin Tg pigs were 11.8% (6/51) and 18.2% (2/11), respectively. In all of the Tg pigs examined by real-time PCR analysis, tissue-specific expression of the transgene was confirmed (type I collagen: skin, tendon, vessels, genitalia; albumin: liver). The production of human proteins derived from BAC transgenes was also confirmed. Fluorescence in situ hybridization analysis indicated that the BAC transgenes transferred into porcine oocytes by ICSI-MGT were integrated into single or multiple sites on the host chromosomes. These data demonstrate that Tg pigs expressing human proteins in a tissue-specific manner can be created using a BAC transgenic construct and the ICSI-MGT method.

Keywords

Transgenic pig ICSI-mediated gene transfer Bacterial artificial chromosome Human protein production 

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Masahito Watanabe
    • 1
    • 2
  • Mayuko Kurome
    • 3
  • Hitomi Matsunari
    • 2
  • Kazuaki Nakano
    • 2
  • Kazuhiro Umeyema
    • 2
    • 4
  • Akira Shiota
    • 5
  • Hiromitsu Nakauchi
    • 1
    • 6
  • Hiroshi Nagashima
    • 1
    • 2
    • 4
  1. 1.Nakauchi Stem Cell and Organ Regeneration ProjectJapan Science and Technology Agency (JST), ERATOTokyoJapan
  2. 2.Department of Life Sciences, School of AgricultureMeiji UniversityKanagawaJapan
  3. 3.Institute of Molecular Animal Breeding and BiotechnologyLudwig-Maximilians-University MunichOberschleissheimGermany
  4. 4.International Cluster for Bio-Resource ResearchMeiji UniversityKanagawaJapan
  5. 5.PhoenixBio Co.HiroshimaJapan
  6. 6.Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical ScienceTokyo UniversityTokyoJapan

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