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

• Published in: Frontiers in Marine Science
• Main Authors: Jin, Kaidi, Zhang, Baolu, Jin, Qianqian, Cai, Zhongqiang, Wei, Lei, Wang, Xiaomei, Zheng, Yanxin, Huang, Baoyu, Zhang, Meiwei, Qi, Yitao, Liu, Yaqiong, Wang, Xiaotong
• Format: Article in Journal/Newspaper
• Language: unknown
• Published: Frontiers Media SA 2021
• Subjects: Pacific; Pacific oyster
• Online Access: http://dx.doi.org/10.3389/fmars.2021.763470; https://www.frontiersin.org/articles/10.3389/fmars.2021.763470/full

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.