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Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases
Author(s) -
Kasparek Petr,
Krausova Michaela,
Haneckova Radka,
Kriz Vitezslav,
Zbodakova Olga,
Korinek Vladimir,
Sedlacek Radislav
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2014.09.014
Subject(s) - transcription activator like effector nuclease , zinc finger nuclease , gene targeting , homologous recombination , genome editing , biology , transgene , crispr , microbiology and biotechnology , gene , embryonic stem cell , genetics
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.