CRISPR/Cas9-Mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda.
CRISPR/Cas9-Mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda.
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CRISPR/Cas9 介导的表白虾 EcChi4 基因组编辑和诱变
DOI：
10.1534/g3.116.034082
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发表时间：
2016-11-08
期刊：
影响因子：
--
通讯作者：
Xiang J
中科院分区：
文献类型：
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作者：
Gui T;Zhang J;Song F;Sun Y;Xie S;Yu K;Xiang J
II型成簇规则间隔短回文重复序列（CRISPR）系统的发展引发了基因工程的革命，该技术已应用于多种物种的基因组编辑。然而，目前还没有关于虾的目标特异性基因组编辑的报道。在本研究中，我们开发了脊尾白虾的显微注射方法，并成功地将CRISPR/Cas9技术应用于脊尾白虾的基因组编辑。通过共注射 Cas9 核酸酶的 mRNA 和专门针对 E. carinicauda 几丁质酶 4 (EcChi4) 的 gRNA，获得了具有 indel 突变的虾。进一步分析表明，这些突变可以遗传给下一代。这是首次在十足目动物中成功演示位点特异性基因组编辑，并将进一步为十足目动物功能基因组学的研究做出贡献。
The development of the type II clustered regularly interspaced short palindromic repeats (CRISPR) system has resulted in the revolution of genetic engineering, and this technology has been applied in the genome editing of various species. However, there are no reports about target-specific genome editing in shrimp. In this research, we developed a microinjection method for the ridgetail white prawn Exopalaemon carinicauda and successfully applied CRISPR/Cas9 technology to the genome editing of E. carinicauda. Through coinjection of mRNA of Cas9 nuclease and gRNA specialized for E. carinicauda chitinase 4 (EcChi4), shrimps with indel mutations were obtained. Further analysis showed that the mutations could be transmitted to the next generation. This is the first time that site-specific genome editing has been successfully demonstrated in a decapod, and will further contribute to the study of functional genomics in decapods.