Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases

0440940 - ÚMG 2015 RIV NL eng J - Journal Article
Kašpárek, Petr - Krausová, Michaela - Hanečková, Radka - Kříž, Vítězslav - Žbodáková, Olga - Kořínek, Vladimír - Sedláček, Radislav
Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases.
FEBS Letters. Roč. 588, č. 21 (2014), 3982-3988. ISSN 0014-5793. E-ISSN 1873-3468
R&D Projects: GA MŠMT(CZ) ED1.1.00/02.0109; GA ČR(CZ) GA14-33952S; GA MŠMT(CZ) LM2011032
Institutional support: RVO:68378050
Keywords : Gene targeting * Homologous recombination * Rosa26 locus * Transgenic * Transcription activator-like effector nucleases * Zinc-finger nucleases
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 3.169, year: 2014

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.
Permanent Link: http://hdl.handle.net/11104/0244021