Efficient gene targeting by TAL effector nucleases coinjected with exonucleases in zygotes.

Efficient gene targeting by TAL effector nucleases coinjected with exonucleases in zygotes.
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DOI:
10.1038/srep01253
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发表时间:
2013
期刊:
影响因子:
4.6
通讯作者:
Serikawa, Tadao
Serikawa, Tadao
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Mashimo, Tomoji;Kaneko, Takehito;Sakuma, Tetsushi;Kobayashi, Junya;Kunihiro, Yayoi;Voigt, Birger;Yamamoto, Takashi;Serikawa, Tadao

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转录激活因子样效应物核酸酶(TALENs)是在多种物种中进行靶向基因编辑的通用工具。然而,它们的效率仍然不足,尤其是在哺乳动物胚胎中。在此,我们表明核酸外切酶1(Exo1)与工程化的位点特异性TALENs共同表达,可在大鼠成纤维细胞中高效破坏内源基因。在受精的大鼠卵子中以及在生产白化(Tyr)基因敲除大鼠时,也观察到Exo1使效率提高了类似的约30%。这些发现表明,TALENs与Exo1联用是一种利用受精卵产生基因敲除的简便且高效的方法,这增加了适用于各种物种的基因靶向技术的范围。
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.
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