Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Sci Rep. 2013;3:1253. doi: 10.1038/srep01253. Epub 2013 Feb 13.
TAL Effector Nucleases (TALENs) are versatile tools for targeted gene editing in various species. However, their efficiency is still insufficient, especially in mammalian embryos. Here, we showed that combined expression of Exonuclease 1 (Exo1) with engineered site-specific TALENs provided highly efficient disruption of the endogenous gene in rat fibroblast cells. A similar increased efficiency of up to ~30% with Exo1 was also observed in fertilized rat eggs, and in the production of knockout rats for the albino (Tyr) gene. These findings demonstrate TALENs with Exo1 is an easy and efficient method of generating gene knockouts using zygotes, which increases the range of gene targeting technologies available to various species.
TAL 效应物核酸酶 (TALENs) 是一种在各种物种中进行靶向基因编辑的多功能工具。然而，其效率仍然不足，尤其是在哺乳动物胚胎中。在这里，我们表明，与工程化的位点特异性 TALENs 联合表达外切核酸酶 1 (Exo1) 可在大鼠成纤维细胞中高效破坏内源性基因。在受精的大鼠卵和 albino (Tyr) 基因敲除大鼠的产生中，也观察到 Exo1 使效率提高了相似的高达~30%。这些发现表明，TALENs 与 Exo1 联合使用是一种使用受精卵生成基因敲除的简便高效方法，这增加了各种物种可用的基因靶向技术的范围。
