Efficient generation of [alpha](1,3) galactosyltransferase knockout porcine fetal fibroblasts for nuclear transfer
Date
2002
Authors
Harrison, S.
Guidolin, A.
Faast, R.
Crocker, L.
Giannakis, C.
d'Apice, A.
Nottle, M.
Lyons, I.
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Journal article
Citation
Transgenic Research, 2002; 11(2):143-150
Statement of Responsibility
Sharon J. Harrison, Angelo Guidolin, Renate Faast, Lesley A. Crocker, Chris Giannakis, Anthony J.F. d’Apice, Mark B. Nottle and Ian Lyons
Conference Name
Abstract
Pigs are currently considered themost likely source of organs for human xenotransplantation because of anatomical and physiological similarities to humans, and the relative ease with which they can be bred in large numbers. A severe form of rejection known as hyperacute rejection has been the major barrier to the use of xenografts. Generating transgenic pigs for organ transplantation is likely to involve precise genetic manipulation to ablate the α(1,3) galactosyltransferase (galT) gene. In contrast to the mouse, homologous recombination in livestock species to ablate genes is hampered by the inability to isolate functional embryonic stem cells. However, nuclear transfer using genetically targeted cultured somatic cells provides an alternativemeans to producing pigs deficient for galT. In this study we successfully produced galT+/− somatic porcine fetal fibroblasts using two approaches; positive negative selection (PNS) using an isogenic targeting construct, and with a promoterless vector using non-isogenic DNA.
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The original publication is available at www.springerlink.com