Production of Genetically Modified Porcine Blastocysts by Somatic Cell Nuclear Transfer: Preliminary Results Toward Production of Xenograft-Competent Miniature Pigs
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概要
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Galα1-3Gal (α-Gal epitope) is the major xenoantigenic epitope responsible for hyperacute rejection upon pig-to-human xenotransplantation. Endo-β-galactosidase C (EndoGalC) from Clostridium perfringens can digest the α-Gal epitope. In this study, gene-engineered primary cultured porcine embryonic fibroblasts (PEF) expressing EndoGalC were obtained and subjected to somatic cell nuclear transfer (SCNT) to test whether xenograft-competent pigs can be created. The EndoGalC-expressing PEF clones exhibited highly reduced expression of α-Gal epitope, as revealed by cytochemical staining with BS-I-B4 isolectin, a lectin that specifically binds to α-Gal epitope, and FACS analysis. The pattern of low level of α-Gal epitope expression continued for at least 6 months (more than 10 generations) after isolation. SCNT of nuclei from these cells resulted in the generation of blastocysts that displayed nearly complete loss of α-Gal epitope from their cell surface. This is the first study to demonstrate that SCNT using EndoGalC-expressing PEFs as donors would be useful for production of genetically modified cloned pigs suitable for xenotransplantation.
著者
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YOSHIDA Mitsutoshi
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University
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MIYOSHI Kazuchika
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University
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HIMAKI Takehiro
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University
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SATO Masahiro
Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University
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WATANABE Satoshi
Animal Genome Research Unit, Division of Animal Science, National Institute of Agrobiological Scienc
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CHI Haiei
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University
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Miyoshi Kazuchika
Laboratory Of Animal Reproduction Faculty Of Agriculture Kagoshima University
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Yoshida Mitsutoshi
Laboratory Of Animal Reproduction Faculty Of Agriculture Kagoshima University
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Himaki Takehiro
Laboratory Of Animal Reproduction Faculty Of Agriculture Kagoshima University
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Sato Masahiro
Section Of Gene Expression Regulation Frontier Science Research Center Kagoshima University
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Watanabe Satoshi
Animal Genome Research Unit Division Of Animal Science National Institute Of Agrobiological Sciences
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Chi Haiei
Laboratory Of Animal Reproduction Faculty Of Agriculture Kagoshima University
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