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Production of gene-targeted sheep by nuclear transfer from cultured somatic cells

Nature volume 405, pages 1066–1069 (2000)Cite this article

An Erratum to this article was published on 02 November 2000

Abstract

It is over a decade since the first demonstration that mouse embryonic stem cells could be used to transfer a predetermined genetic modification to a whole animal1. The extension of this technique to other mammalian species, particularly livestock, might bring numerous biomedical benefits, for example, ablation of xenoreactive transplantation antigens, inactivation of genes responsible for neuropathogenic disease and precise placement of transgenes designed to produce proteins for human therapy. Gene targeting has not yet been achieved in mammals other than mice, however, because functional embryonic stem cells have not been derived. Nuclear transfer from cultured somatic cells provides an alternative means of cell-mediated transgenesis2,3. Here we describe efficient and reproducible gene targeting in fetal fibroblasts to place a therapeutic transgene at the ovine α1(I) procollagen (COL1A1) locus and the production of live sheep by nuclear transfer.

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Figure 1: Diagrams of ovine COL1A1, COLT-1 and COLT-2 targeting vectors and targeted COL1A1 locus.
Figure 2: Analysis of COLT-2 transfected (PDCAAT) cell clones.
Figure 3: Gene-targeted lambs.
Figure 4: Southern analysis of nuclear transfer lambs.

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Acknowledgements

We would like to acknowledge the contributions of Y. Gibson and K. Mycock for embryo manipulation; E. Emslie and L. Hutchison for molecular biology technical assistance; T. Johnston for protein analysis; and the PPL Therapeutics large animal team for animal husbandry and veterinary procedures. We thank I. Garner and D. Ayares for useful discussions.

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Author notes

  1. K. H. S. Campbell

    Present address: Division of Animal Physiology, University of Nottingham, School of Biological Sciences, Loughborough, LE12 5RD, UK

  2. Correspondence and requests for materials should be addressed to A.J.K.

Authors and Affiliations

  1. PPL Therapeutics Ltd, Roslin, Edinburgh, EH25 9PP, UK

    K. J. McCreath, J. Howcroft, K. H. S. Campbell, A. Colman, A. E. Schnieke & A.J. Kind

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  1. K. J. McCreath
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Correspondence to A.J. Kind.

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McCreath, K., Howcroft, J., Campbell, K. et al. Production of gene-targeted sheep by nuclear transfer from cultured somatic cells. Nature 405, 1066–1069 (2000). https://doi.org/10.1038/35016604

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