Application of oocyte cryopreservation technology in TALEN-mediated mouse genome editing.
Application of oocyte cryopreservation technology in TALEN-mediated mouse genome editing.
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DOI：
10.1538/expanim.63.349
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发表时间：
2014
影响因子：
2.4
通讯作者：
Yamamoto T
中科院分区：
文献类型：
--
作者：
Nakagawa Y;Sakuma T;Nakagata N;Yamasaki S;Takeda N;Ohmuraya M;Yamamoto T
生殖工程技术，如体外受精（IVF）和胚胎或精子的冷冻保存，对于基因工程小鼠的保存、繁殖和运输是必不可少的。然而，目前尚不清楚这些技术是否可以应用于使用工程核酸酶（如转录激活因子样效应核酸酶（TALENs））产生基因组编辑小鼠。在这里，我们证明了使用冷冻精子在体外受精的冷冻卵母细胞对小鼠进行talen介导的基因组编辑的有效性。我们检查了用于TALEN注射的卵母细胞来源的精子（新鲜与冷冻）、受精方法（交配与体外受精）和受精卵母细胞（新鲜与冷冻）的并排比较；我们发现在所有测试条件下产生的受精卵母细胞都适用于talen介导的突变。此外，我们通过比较5周龄和8 - 12周龄的母鼠作为TALEN注射卵母细胞的来源，研究了母鼠的周龄是否会影响基因修饰的效率。当5周龄或8 - 12周龄的小鼠使用或不冷冻卵母细胞时，内源性基因的基因组编辑效率始终为95-100%。因此，我们的报告描述了冻融卵母细胞和来自不同周龄雌性小鼠的卵母细胞用于talen介导的基因组编辑的可用性，从而提高了这种创新基因靶向策略的便利性。
Reproductive engineering techniques, such as in vitro fertilization (IVF) and cryopreservation of embryos or spermatozoa, are essential for preservation, reproduction, and transportation of genetically engineered mice. However, it has not yet been elucidated whether these techniques can be applied for the generation of genome-edited mice using engineered nucleases such as transcription activator-like effector nucleases (TALENs). Here, we demonstrate the usefulness of frozen oocytes fertilized in vitro using frozen sperm for TALEN-mediated genome editing in mice. We examined side-by-side comparisons concerning sperm (fresh vs. frozen), fertilization method (mating vs. IVF), and fertilized oocytes (fresh vs. frozen) for the source of oocytes used for TALEN injection; we found that fertilized oocytes created under all tested conditions were applicable for TALEN-mediated mutagenesis. In addition, we investigated whether the ages in weeks of parental female mice can affect the efficiency of gene modification, by comparing 5-week-old and 8–12-week-old mice as the source of oocytes used for TALEN injection. The genome editing efficiency of an endogenous gene was consistently 95–100% when either 5-week-old or 8–12-week-old mice were used with or without freezing the oocytes. Thus, our report describes the availability of freeze-thawed oocytes and oocytes from female mice at various weeks of age for TALEN-mediated genome editing, thus boosting the convenience of such innovative gene targeting strategies.
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影响因子：
4.6
作者：
Sakuma, Tetsushi;Ochiai, Hiroshi;Kaneko, Takehito;Mashimo, Tomoji;Tokumasu, Daisuke;Sakane, Yuto;Suzuki, Ken-ichi;Miyamoto, Tatsuo;Sakamoto, Naoaki;Matsuura, Shinya;Yamamoto, Takashi
通讯作者：
Yamamoto, Takashi
DOI：
10.1073/pnas.1218721110
发表时间：
2013-03-05
影响因子：
11.1
作者：
Wefers, Benedikt;Meyer, Melanie;Kuehn, Ralf
通讯作者：
Kuehn, Ralf
影响因子：
3.3
作者：
Panda, Sudeepta Kumar;Wefers, Benedikt;Kuehn, Ralf
通讯作者：
Kuehn, Ralf
影响因子：
2.7
作者：
Ma, Ke;Wang, Jianying;Li, Zheng
通讯作者：
Li, Zheng
通过类转录激活因子效应核酸酶在小鼠中实现高效、可遗传的基因靶向
影响因子：
14.9
作者：
Qiu Z;Liu M;Chen Z;Shao Y;Pan H;Wei G;Yu C;Zhang L;Li X;Wang P;Fan HY;Du B;Liu B;Liu M;Li D
通讯作者：
Li D