Abstract
Nuclear transfer offers a cell-based route for producing precise genetic modifications in a range of animal species. Using sheep, we report reproducible targeted gene deletion at two independent loci in fetal fibro-blasts. Vital regions were deleted from the α(1,3)galactosyl transferase (GGTA1) gene, which may account for the hyperacute rejection of xenografted organs, and from the prion protein (PrP) gene, which is directly associated with spongiform encephalopathies in humans and animals. Reconstructed embryos were prepared using cultures of targeted or nontargeted donor cells. Eight pregnancies were maintained to term and four PrP−/+ lambs were born. Although three of these perished soon after birth, one survived for 12 days. These data show that lambs carrying targeted gene deletions can be generated by nuclear transfer.
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Acknowledgements
The authors would like to thank J. Bowering, W. Bosma, P. Johnson, T. Ferrier, D. McGavin, B. Gasparrini, and L. Harkness for technical support, and Jane Lebkowski for reading the manuscript. The Biotechnology and Biological Sciences Research Council and the Geron Corporation provided financial support.
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Denning, C., Burl, S., Ainslie, A. et al. Deletion of the α(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. Nat Biotechnol 19, 559–562 (2001). https://doi.org/10.1038/89313
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DOI: https://doi.org/10.1038/89313
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