Universität zu Köln

Sommer, Daniel, Peters, Annika, Wirtz, Tristan, Mai, Maren, Ackermann, Justus, Thabet, Yasser, Schmidt, Juergen, Weighardt, Heike, Wunderlich, F. Thomas, Degen, Joachim, Schultze, Joachim L. and Beyer, Marc ORCID: 0000-0001-9704-148X (2014). Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases. Nat. Commun., 5. LONDON: NATURE PUBLISHING GROUP. ISSN 2041-1723

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Sommer, Daniel UNSPECIFIED UNSPECIFIED UNSPECIFIED Peters, Annika UNSPECIFIED UNSPECIFIED UNSPECIFIED Wirtz, Tristan UNSPECIFIED UNSPECIFIED UNSPECIFIED Mai, Maren UNSPECIFIED UNSPECIFIED UNSPECIFIED Ackermann, Justus UNSPECIFIED UNSPECIFIED UNSPECIFIED Thabet, Yasser UNSPECIFIED UNSPECIFIED UNSPECIFIED Schmidt, Juergen UNSPECIFIED UNSPECIFIED UNSPECIFIED Weighardt, Heike UNSPECIFIED UNSPECIFIED UNSPECIFIED Wunderlich, F. Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Degen, Joachim UNSPECIFIED UNSPECIFIED UNSPECIFIED Schultze, Joachim L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Beyer, Marc UNSPECIFIED orcid.org/0000-0001-9704-148X UNSPECIFIED
URN:	urn:nbn:de:hbz:38-452571
DOI:	10.1038/ncomms4045
Journal or Publication Title:	Nat. Commun.
Volume:	5
Date:	2014
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	2041-1723
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ZINC-FINGER NUCLEASES; ONE-STEP GENERATION; MULTIPLE GENES; KNOCKOUT RATS; TALENS; SATB1; MICROINJECTION; SPECIFICITY; EXPRESSION; ZEBRAFISH Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/45257