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Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases
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SYSNO ASEP 0440940 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases Author(s) Kašpárek, Petr (UMG-J)
Krausová, Michaela (UMG-J) RID
Hanečková, Radka (UMG-J)
Kříž, Vítězslav (UMG-J)
Žbodáková, Olga (UMG-J)
Kořínek, Vladimír (UMG-J) RID
Sedláček, Radislav (UMG-J) RIDSource Title FEBS Letters. - : Wiley - ISSN 0014-5793
Roč. 588, č. 21 (2014), 3982-3988Number of pages 7 s. Language eng - English Country NL - Netherlands Keywords Gene targeting ; Homologous recombination ; Rosa26 locus ; Transgenic ; Transcription activator-like effector nucleases ; Zinc-finger nucleases Subject RIV EB - Genetics ; Molecular Biology R&D Projects ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA14-33952S GA ČR - Czech Science Foundation (CSF) LM2011032 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UMG-J - RVO:68378050 UT WOS 000344075400029 DOI 10.1016/j.febslet.2014.09.014 Annotation Gene targeting in mice mainly employs homologous recombination (HR) in embryonic stem (ES) cells. Although it is a standard way for production of genetically modified mice, the procedure is laborious and time-consuming. This study describes targeting of the mouse Rosa26 locus by transcription activator-like effector nucleases (TALENs). We employed TALEN-assisted HR in zygotes to introduce constructs encoding TurboRFP and TagBFP fluorescent proteins into the first intron of the Rosa26 gene, and in this way generated two transgenic mice. We also demonstrated that these Rosa26-specific TALENs exhibit high targeting efficiency superior to that of zinc-finger nucleases (ZFNs) specific for the same targeting sequence. Moreover, we devised a reporter assay to assess TALENs activity and specificity to improve the quality of TALEN-assisted targeting. (C) 2014 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2015
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