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

Daniel Sommer, Annika E. Peters, Tristan Wirtz, Maren Mai, Justus Ackermann, Yasser Thabet, Jürgen Schmidt, Heike Weighardt, F. Thomas Wunderlich, Joachim Degen, Joachim L. Schultze and Marc Beyer ()
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Daniel Sommer: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Annika E. Peters: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Tristan Wirtz: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Maren Mai: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Justus Ackermann: Max Planck Institute for Neurological Research and Institute for Genetics, University of Cologne
Yasser Thabet: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Jürgen Schmidt: University Hospital Bonn
Heike Weighardt: Immunology and Environment, LIMES Institute, University of Bonn
F. Thomas Wunderlich: Max Planck Institute for Neurological Research and Institute for Genetics, University of Cologne
Joachim Degen: Genomic Resource Centre, LIMES Institute, University of Bonn
Joachim L. Schultze: Genomics and Immunoregulation, LIMES Institute, University of Bonn
Marc Beyer: Genomics and Immunoregulation, LIMES Institute, University of Bonn

Nature Communications, 2014, vol. 5, issue 1, 1-12

Abstract: 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.

Date: 2014
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DOI: 10.1038/ncomms4045

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