Targeted disruption of the α1,3-galactosyltransferase gene in cloned pigs (original) (raw)
- Article
- Published: 01 March 2002
- Todd D. Vaught1,
- Jeremy Boone1,
- Shu-Hung Chen1,
- Carol J. Phelps1,
- Suyapa Ball1,
- Jeff A. Monahan1,
- Peter M. Jobst1,
- Kenneth J. McCreath2,
- Ashley E. Lamborn1,
- Jamie L. Cowell-Lucero1,
- Kevin D. Wells1,
- Alan Colman2,
- Irina A. Polejaeva1 &
- …
- David L. Ayares1
Nature Biotechnology volume 20, pages 251–255 (2002)Cite this article
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Abstract
Galactose-α1,3-galactose (α1,3Gal) is the major xenoantigen causing hyperacute rejection in pig-to-human xenotransplantation. Disruption of the gene encoding pig α1,3-galactosyltransferase (α1,3GT) by homologous recombination is a means to completely remove the α1,3Gal epitopes from xenografts. Here we report the disruption of one allele of the pig α1,3GT gene in both male and female porcine primary fetal fibroblasts. Targeting was confirmed in 17 colonies by Southern blot analysis, and 7 of them were used for nuclear transfer. Using cells from one colony, we produced six cloned female piglets, of which five were of normal weight and apparently healthy. Southern blot analysis confirmed that these five piglets contain one disrupted pig α1,3GT allele.
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Acknowledgements
We thank B. Gragg, W. Lucero, T. Akers, H. Bishop, and J. McPherson for technical contributions to embryo transfer and animal husbandry; A. Garst and J. Hencke for help in nuclear transfer; C. Koike and T. E. Starzl at the University of Pittsburgh for providing us with porcine α1,3GT gene genomic sequence; M. Moore, A. Kind, and A. Schnieke for valuable contributions in discussions; and staff at the Virginia Maryland Regional Veterinary College for prenatal care of some of the nuclear transfer recipients and physical examination of piglets. This research was funded in part by a grant from the National Institute of Standards and Technology (NIST) Advanced Technology Program (ATP).
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Authors and Affiliations
- PPL Therapeutics Inc., 1700 Kraft Drive, Blacksburg, 24060, Virginia, USA
Yifan Dai, Todd D. Vaught, Jeremy Boone, Shu-Hung Chen, Carol J. Phelps, Suyapa Ball, Jeff A. Monahan, Peter M. Jobst, Ashley E. Lamborn, Jamie L. Cowell-Lucero, Kevin D. Wells, Irina A. Polejaeva & David L. Ayares - PPL Therapeutics Ltd., Roslin, EH25 9PP, Midlothian, UK
Kenneth J. McCreath & Alan Colman
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- Yifan Dai
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Correspondence toYifan Dai.
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Dai, Y., Vaught, T., Boone, J. et al. Targeted disruption of the α1,3-galactosyltransferase gene in cloned pigs.Nat Biotechnol 20, 251–255 (2002). https://doi.org/10.1038/nbt0302-251
- Received: 08 January 2002
- Accepted: 22 January 2002
- Issue Date: 01 March 2002
- DOI: https://doi.org/10.1038/nbt0302-251