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Glycobiology Advance Access originally published online on October 17, 2007
Glycobiology 2008 18(1):9-19; doi:10.1093/glycob/cwm114
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Production and Characterization of Transgenic Mice Systemically Expressing Endo-β-Galactosidase C

Satoshi Watanabe1,2, Masako Misawa3, Takashi Matsuzaki3,4, Takayuki Sakurai5, Takashi Muramatsu6, Taka-aki Yokomine7 and Masahiro Sato7

2 Animal Genome Research Unit, Division of Animal Science, National Institute of Agrobiological Sciences, 2 Ikenodai, Tsukuba, Ibaraki, 305-0901, Japan
3 Division of Resources Life Science, United Graduate School of Agricultural Sciences, Tottori University, 4-101 Koyama-Minami, Tottori, 680-8553, Japan
4 Department of Biological Science, Faculty of life and Environmental Science, Shimane University, 1060 Nishikawatsu-cho, Matsue, Shimane, 690-8504, Japan
5 Department of Organ Regeneration, Graduate School of Medicine, Shinshu University 3-1-1 Asahi, Matsumoto 390-8621, Japan
6 Department of Health Science, Faculty of Psychological and Physical Sciences, Aichi Gakuin University, 12 Araike, Iwasaki-cho, Nisshin, Aichi, 470-0195, Japan
7 Frontier Science Research Center, Kagoshima University, Korimoto 1-21-40, Kagoshima 890-0065, Japan

1 To whom correspondence should be addressed: Tel and Fax: +81-29-838-8662 e-mail: kettle{at}affrc.go.jp

Received on March 1, 2007; revised on September 26, 2007; accepted on October 5, 2007

The {alpha}Gal epitope (Gal{alpha}1-3Gal) is a sugar structure expressed on the cell surface of almost all organisms except humans and old-world-monkeys, which express natural anti-{alpha}Gal antibodies. The presence of these antibodies elicits a hyper acute rejection (HAR) upon xenotransplantation of cellular materials, such as from pigs to human beings. Endo-β-galactosidase C (EndoGalC), an enzyme isolated from Clostridium perfringens, removes the {alpha}Gal epitope by cleaving the Galβ1-4GlcNAc linkage in the Gal{alpha}1-3Galβ1-4GlcNAc sequence. To explore the possibility that cells or organs from transgenic pigs systemically expressing EndoGalC might be suitable for xenotransplantation, we first introduced the EndoGalC transgene into the mouse genome via pronuclear injection. The progeny of the resulting transgenics expressed EndoGalC mRNA and protein. Flow cytometry and histochemical analyses revealed a dramatic reduction in the expression of the {alpha}Gal epitope in these mice. They also exhibited abnormal phenotypes, such as occasional death immediately after birth, growth retardation, and transient skin lesions. Interestingly, the phenotypic abnormalities seen in these transgenics were similar to those observed in β1,4-galactosyltransferase 1 (β4GalT-1) knockout (KO) mice. Most probably, these phenotypes were caused by exposure of the internal N-acetylglucosamine residue at the end of the sugar chain on the cell surface. The present findings also provide some basis for evaluating possible application of the transgenic approach for xenotranplantation.

Key words: {alpha}Gal epitope / galactosidase lectin / systemic promoter / transgenic mice


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