Somatic cell reprogramming-free generation of genetically modified pigs.
Somatic cell reprogramming-free generation of genetically modified pigs.
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DOI：
10.1126/sciadv.1600803
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发表时间：
2016-09
期刊：
影响因子：
13.6
通讯作者：
Otoi T
中科院分区：
文献类型：
--
作者：
Tanihara F;Takemoto T;Kitagawa E;Rao S;Do LT;Onishi A;Yamashita Y;Kosugi C;Suzuki H;Sembon S;Suzuki S;Nakai M;Hashimoto M;Yasue A;Matsuhisa M;Noji S;Fujimura T;Fuchimoto D;Otoi T
一种通过将CRISPR/Cas9系统电穿孔到合子中来产生突变猪的新的高效方法。用于生物医学应用的转基因猪主要是使用体细胞核移植技术产生的;然而，这种方法需要复杂的显微操作技术，并且有时会通过供体体细胞核的错误表观遗传重编程增加产前和产后死亡的风险。因此，转基因猪的生产并没有得到广泛应用。我们提供了一种简单的方法，用于在猪中进行CRISPR（成簇的规则间隔短回文重复序列）/Cas9基因编辑，该方法包括通过电穿孔将Cas9蛋白和单向导RNA引入体外受精卵中。通过电穿孔Cas9蛋白（GEEP）进行基因编辑的使用导致高效的靶向基因破坏，并通过肌生长抑制素突变猪的有效生产得到验证。由于GEEP不需要与用于体细胞重编程的显微操作相关的复杂方法，因此它具有促进猪的遗传修饰的潜力。
A new and highly efficient method for generating mutant pigs by electroporating the CRISPR/Cas9 system into zygotes. Genetically modified pigs for biomedical applications have been mainly generated using the somatic cell nuclear transfer technique; however, this approach requires complex micromanipulation techniques and sometimes increases the risks of both prenatal and postnatal death by faulty epigenetic reprogramming of a donor somatic cell nucleus. As a result, the production of genetically modified pigs has not been widely applied. We provide a simple method for CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 gene editing in pigs that involves the introduction of Cas9 protein and single-guide RNA into in vitro fertilized zygotes by electroporation. The use of gene editing by electroporation of Cas9 protein (GEEP) resulted in highly efficient targeted gene disruption and was validated by the efficient production of Myostatin mutant pigs. Because GEEP does not require the complex methods associated with micromanipulation for somatic reprogramming, it has the potential for facilitating the genetic modification of pigs.
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影响因子：
4.6
作者：
Hashimoto M;Takemoto T
通讯作者：
Takemoto T
影响因子：
56.9
作者：
Onishi, A;Iwamoto, M;Perry, ACF
通讯作者：
Perry, ACF
影响因子：
3
作者：
Tan W;Proudfoot C;Lillico SG;Whitelaw CB
通讯作者：
Whitelaw CB
影响因子：
2.6
作者：
Kues, Wilfried A.;Niemann, Heiner
通讯作者：
Niemann, Heiner
影响因子：
30.8
作者：
Sekine, K;Ohuchi, H;Kato, S
通讯作者：
Kato, S