Efficient gene knockout in goats using CRISPR/Cas9 system.

Efficient gene knockout in goats using CRISPR/Cas9 system.
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使用 CRISPR/Cas9 系统有效敲除山羊基因

DOI:
10.1371/journal.pone.0106718
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发表时间:
2014
期刊:
影响因子:
3.7
通讯作者:
Chen C
Chen C
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Ni W;Qiao J;Hu S;Zhao X;Regouski M;Yang M;Polejaeva IA;Chen C

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CRISPR/Cas9系统已被用作实验室动物(如小鼠、大鼠、斑马鱼和猪)的有效基因组编辑工具。在这里,我们报告了CRISPR/Cas9介导的方法可以有效地诱导山羊原代成纤维细胞中的单等位基因和双等位基因基因敲除。通过CRISPR/Cas9介导的基因组编辑,在山羊成纤维细胞中同时破坏了四个基因。单基因敲除的成纤维细胞被成功地用于体细胞核移植(SCNT),并导致活产山羊携带双等位基因突变。CRISPR/Cas9系统代表了一种高效、简便的大型动物基因组靶向编辑平台,可广泛应用于生物医学和农业应用。
The CRISPR/Cas9 system has been adapted as an efficient genome editing tool in laboratory animals such as mice, rats, zebrafish and pigs. Here, we report that CRISPR/Cas9 mediated approach can efficiently induce monoallelic and biallelic gene knockout in goat primary fibroblasts. Four genes were disrupted simultaneously in goat fibroblasts by CRISPR/Cas9-mediated genome editing. The single-gene knockout fibroblasts were successfully used for somatic cell nuclear transfer (SCNT) and resulted in live-born goats harboring biallelic mutations. The CRISPR/Cas9 system represents a highly effective and facile platform for targeted editing of large animal genomes, which can be broadly applied to both biomedical and agricultural applications.
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