AI Article Synopsis
- Gene targeting in mice typically uses homologous recombination in embryonic stem cells, but this process is time-consuming and complex.
- This study introduces a method using transcription activator-like effector nucleases (TALENs) to target the Rosa26 gene locus, successfully creating two transgenic mice with fluorescent proteins.
- The research shows that the TALENs used have greater targeting efficiency than zinc-finger nucleases, and a new reporter assay was developed to evaluate TALENs' effectiveness and precision.
Article Abstract
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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| http://dx.doi.org/10.1016/j.febslet.2014.09.014 | DOI Listing |
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