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

First Author  Sommer D Year  2014
Journal  Nat Commun Volume  5
Pages  3045 PubMed ID  24413636
Mgi Jnum  J:206457 Mgi Id  MGI:5550309
Doi  10.1038/ncomms4045 Citation  Sommer D, et al. (2014) Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases. Nat Commun 5:3045
abstractText  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.
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