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Generation of TALEN-mediated GRdim knock-in rats by homologous recombination.

ABSTRACT: Transcription Activator-Like Effector Nucleases (TALEN) are potential tools for precise genome engineering of laboratory animals. We report the first targeted genomic integration in the rat using TALENs (Transcription Activator-Like Effector Nucleases) by homology-derived recombination (HDR). We assembled TALENs and designed a linear donor insert targeting a pA476T mutation in the rat Glucocorticoid Receptor (Nr3c1) namely GR(dim), that prevents receptor homodimerization in the mouse. TALEN mRNA and linear double-stranded donor were microinjected into rat one-cell embryos. Overall, we observed targeted genomic modifications in 17% of the offspring, indicating high TALEN cutting efficiency in rat zygotes.

SUBMITTER: Ponce de Leon V

PROVIDER: S-EPMC3921256 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Generation of TALEN-mediated GRdim knock-in rats by homologous recombination.

Ponce de León Verónica V Mérillat Anne-Marie AM Tesson Laurent L Anegón Ignacio I Hummler Edith E

PloS one 20140211 2

Transcription Activator-Like Effector Nucleases (TALEN) are potential tools for precise genome engineering of laboratory animals. We report the first targeted genomic integration in the rat using TALENs (Transcription Activator-Like Effector Nucleases) by homology-derived recombination (HDR). We assembled TALENs and designed a linear donor insert targeting a pA476T mutation in the rat Glucocorticoid Receptor (Nr3c1) namely GR(dim), that prevents receptor homodimerization in the mouse. TALEN mRNA ...[more]

PMID: 24523878

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