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CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation

文献类型： 外文期刊

作者： Jin, Kaidi 1 ; Zhang, Baolu 2 ; Jin, Qianqian 1 ; Cai, Zhongqiang 3 ; Wei, Lei 1 ; Wang, Xiaomei 3 ; Zheng, Yanxin 3 ; Hu 1 ;

作者机构： 1.Ludong Univ, Sch Agr, Yantai, Peoples R China

2.Minist Nat Resources Peoples Republ China, Ocean Consultat Ctr, Beijing, Peoples R China

3.Chinese Acad Fishery Sci, Changdao Enhancement & Expt Stn, Yantai, Peoples R China

4.Shaanxi Normal Univ, Natl Engn Lab Resource Developing Endangered Chin, Coll Life Sci, Key Lab,Minist Educ Med Resources & Nat Pharmaceu, Xian, Peoples R China

关键词： CRISPR; Cas9; electroporation; Crassostrea angulate; gene editing; mollusk

期刊名称：FRONTIERS IN MARINE SCIENCE （ 影响因子：4.912； 五年影响因子：5.125 ）

ISSN：

年卷期： 2021 年 8 卷

页码：

收录情况： SCI

摘要： The Fujian oyster (Crassostrea angulate) is an important marine bivalve mollusk with high economic value. Gene function research and gene editing techniques have broad application prospects in oyster. The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been widely used for genome engineering in many species. CRISPR-mediated gene editing has also been used successfully in the Pacific oyster through direct delivery of the CRISPR/Cas9 components into oyster embryos by microinjection. However, the low throughput and operational difficulties associated with microinjection is one of the factors limiting the widespread application of CRISPR/Cas9 in oysters. In this study, we attempted to deliver the CRISPR/Cas9-system into the embryos of C. angulate by electroporation. An all-in-one CRISPR/Cas9 vector plasmid was used as CRISPR/Cas9 system in this study. Electroporation was carried out using both eggs and blastula larvae. A large number of larvae became malformed or die after electroporation. A single base substitution mutation was detected in the D-larvae developed from electroporated eggs. Our results demonstrate that the CRISPR/Cas9 system can be delivered into embryos of C. angulate for gene editing by electroporation, which provides a reference and will further contribute to the future application of electroporation in mollusks.

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