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Nat Commun. 2014;5:3045. doi: 10.1038/ncomms4045.

Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases.

Author information

  • 1Genomics and Immunoregulation, LIMES Institute, University of Bonn, Carl-Troll-Strasse 31, D-53115 Bonn, Germany.
  • 21] Genomics and Immunoregulation, LIMES Institute, University of Bonn, Carl-Troll-Strasse 31, D-53115 Bonn, Germany [2].
  • 3Max Planck Institute for Neurological Research and Institute for Genetics, University of Cologne, Gleuelerstrasse 50, D-50931 Cologne, Germany.
  • 4Department of Experimental Therapy, University Hospital Bonn, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany.
  • 5Immunology and Environment, LIMES Institute, University of Bonn, Carl-Troll-Strasse 31, D-53115 Bonn, Germany.
  • 6Genomic Resource Centre, LIMES Institute, University of Bonn, Carl-Troll-Strasse 31, D-53115 Bonn, Germany.

Abstract

Generation of mouse models by introducing transgenes using homologous recombination is critical for understanding fundamental biology and pathology of human diseases. Here we investigate whether artificial transcription activator-like effector nucleases (TALENs)-powerful tools that induce DNA double-strand breaks at specific genomic locations-can be combined with a targeting vector to induce homologous recombination for the introduction of a transgene in embryonic stem cells and fertilized murine oocytes. We describe the generation of a conditional mouse model using TALENs, which introduce double-strand breaks at the genomic locus of the special AT-rich sequence-binding protein-1 in combination with a large 14.4 kb targeting template vector. We report successful germline transmission of this allele and demonstrate its recombination in primary cells in the presence of Cre-recombinase. These results suggest that TALEN-assisted induction of DNA double-strand breaks can facilitate homologous recombination of complex targeting constructs directly in oocytes.

PMID:
24413636
[PubMed - in process]
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