title: 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.
authors: Department of Pharmacology and Toxicology, University of Lausanne, Lausanne, Switzerland.
authors: Verónica
authors: V
authors: Ponce de León
authors: Department of Pharmacology and Toxicology, University of Lausanne, Lausanne, Switzerland.
authors: Anne-Marie
authors: AM
authors: Mérillat
authors: INSERM UMR 1064 and Transgenic Rat Facility, University of Nantes, Nantes, France.
authors: Laurent
authors: L
authors: Tesson
authors: INSERM UMR 1064 and Transgenic Rat Facility, University of Nantes, Nantes, France.
authors: Ignacio
authors: I
authors: Anegón
authors: Department of Pharmacology and Toxicology, University of Lausanne, Lausanne, Switzerland.
authors: Edith
authors: E
authors: Hummler
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D045744:Cell Line, Tumor; D004720:Endonucleases / Q000235:genetics*; D005260:Female; D055879:Gene Knock-In Techniques; D005818:Genetic Engineering; D005838:Genotype; D059765:Homologous Recombination; D008297:Male; D051379:Mice; D009154:Mutation; D055503:Protein Multimerization; D051381:Rats; D017207:Rats, Sprague-Dawley; D011965:Receptors, Glucocorticoid / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: C542344:NR3C1 protein, rat; D011965:Receptors, Glucocorticoid; D004720:Endonucleases