Genome Editing in the Olive Flounder (Paralichthys olivaceus) Using CRISPR/Cas9 and a Simple Microinjection System

• Published in: Journal of Ocean University of China
• Main Authors: Tan Xungang, Wang Ling, Wu Zhihao, Jiao Shuang, Wang Lijuan, Zou Yuxia, Jiang Jingteng, You Feng
• Format: Report
• Language: English
• Published: OCEAN UNIV CHINA 2021
• Subjects: CRISPR; Cas9; gsdf; microinjection; myomaker; needle with brake; olive flounder; Oceanography; GERM-LINE TRANSMISSION; GENE-TRANSFER; TARGETED MUTAGENESIS; ATLANTIC SALMON; EXPRESSION; EGGS; FISH; NUCLEASE; CELLS; Atlantic salmon
• Online Access: http://ir.qdio.ac.cn/handle/337002/177211; https://doi.org/10.1007/s11802-021-4772-6

The whole-genome sequence of the olive flounder (Paralichthys olivaceus) provides a basis for gene functional analyses, which is important for the aquaculture industry. Understanding gene function will help us to select better economic traits such as fast growth and better culture conditions, which further will increase the aquaculture output. Gene knockout is an important reverse genetics approach for in vivo studies of gene function. In this study, the CRISPR/Cas9 genome editing method with a microinjection system using a simple braked needle was employed in olive flounder. After injection in embryos, green fluorescent protein expression was detected in 40% of larvae. The proportion of normal-hatched larvae was approximately 50%. Different mutations, including short indels and fragment deletions, were found in our test genes gsdf and myomaker. Additionally, we detected more than one mutation in a single larva. In summary, our microinjection technique and CRISPR/Cas9 can be applied to study gene functions in olive flounder.