Generation of TALEN-mediated GRdim knock-in rats by homologous recombination.
basic_science · Level V
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- Record sourced from PubMed, PMID 24523878.
- Also identified by DOI 10.1371/journal.pone.0088146 and PMC identifier 3921256.
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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.
Medical subject headings
- Endonucleases
- Gene Knock-In Techniques
- Homologous Recombination
- Receptors, Glucocorticoid