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Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases.
Kasparek, Petr; Krausova, Michaela; Haneckova, Radka; Kriz, Vitezslav; Zbodakova, Olga; Korinek, Vladimir; Sedlacek, Radislav.
Afiliación
  • Kasparek P; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic.
  • Krausova M; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic.
  • Haneckova R; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic.
  • Kriz V; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic.
  • Zbodakova O; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic.
  • Korinek V; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic. Electronic address: vladimir.korinek@img.cas.cz.
  • Sedlacek R; Institute of Molecular Genetics of the AS CR, v.v.i., Videnska 1083, 142 20, Czech Republic. Electronic address: radislav.sedlacek@img.cas.cz.
FEBS Lett ; 588(21): 3982-8, 2014 Nov 03.
Article en En | MEDLINE | ID: mdl-25241166
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cigoto / Marcación de Gen / Endonucleasas / Sitios Genéticos Límite: Animals Idioma: En Revista: FEBS Lett Año: 2014 Tipo del documento: Article País de afiliación: República Checa Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cigoto / Marcación de Gen / Endonucleasas / Sitios Genéticos Límite: Animals Idioma: En Revista: FEBS Lett Año: 2014 Tipo del documento: Article País de afiliación: República Checa Pais de publicación: Reino Unido