The Derivation of Highly Germline-Competent Embryonic Stem Cells Containing NOD-Derived Genome

  1. Frances A. Brook1,
  2. Edward P. Evans1,
  3. Christopher J. Lord2,
  4. Paul A. Lyons2,
  5. Daniel B. Rainbow2,
  6. Sarah K. Howlett2,
  7. Linda S. Wicker2,
  8. John A. Todd2 and
  9. Richard L. Gardner1
  1. 1Department of Zoology, University of Oxford, Oxford, U.K.
  2. 2JDRF/WT Diabetes and Inflammation Laboratory, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, U.K.

    Abstract

    It would be extremely advantageous to the analysis of disease mechanisms in the spontaneous mouse model of type 1 diabetes, the nonobese diabetic (NOD) strain, if genes in this strain could be modified in vivo using embryonic stem (ES) cells and homologous recombination. However, a NOD ES cell line with adequate germline transmission has not yet been reported. We report the development of highly germline-competent ES cell lines from the F1 hybrid of NOD and 129 for use in NOD gene targeting. Consequently, we developed ES cell lines derived from (NOD × 129)F1 × 129 backcross 1 mice, which were intercrossed to select for homozygosity of particular regions of NOD genome known to contain disease loci.

    Footnotes

    • Address correspondence and reprint requests to Dr. Frances A. Brook, Mammalian Development Laboratory, University of Oxford, Department of Zoology, South Parks Rd., Oxford, OX1 3PS U.K. E-mail: frances.brook{at}zoo.ox.ac.uk.

      Received for publication 31 July 2002 and accepted in revised form 23 September 2002.

      C.J.L. is currently affiliated with Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, London, U.K.

      ES, embryonic stem; LIF, leukemia inhibitory factor.

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