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KMID : 1023620150390030059
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Reproductive and Developmental Biology
2015 Volume.39 No. 3 p.59 ~ p.67
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Knock-in Somatic Cells of Human Decay Accelerating Factor and α1,2-Fucosyltransferase Gene on the α1,3-Galactosyltransferase Gene Locus of Miniature Pig
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Kim Ji-Woo
Kang Man-Jong
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Abstract
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Galactose-α1,3-galactose (α1,3-Gal) epitope is synthesized at a high concentration on the surface of pig cells by α1,3-galactosyltransferase gene (GGTA1). The α1,3-Gal is responsible for hyperacute rejection in pig-to-human xenotransplantation. The generation of transgenic pigs as organ donors for humans is necessary to eliminate the GGTA1 gene that synthesize Galα(1,3)Gal. To prevent hyperacute graft rejection in pig-to-human xenotransplantation, previously, we developed α1,3-galactosyltransferase gene-knock-out somatic cell by homologous recombination. In this study, we established cell lines of α1,3-GT knock-out expressing hDAF and hHT gene from minipig fibroblasts to apply somatic cell nuclear transfer. The hDAF and hHT mRNA were expressed in the knock-in somatic cells and α1,3-GT mRNA was suppressed. However, the knock-in somatic cells were increased resistance to human serum-mediated cytolysis.
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KEYWORD
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Somatic cells, Knock-in, Gene targeting, Xenotransplantation
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