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

• Published in: Journal of Ocean University of China Vol. 20; no. 6; pp. 1528 - 1536
• Main Authors: Tan, Xungang, Wang, Ling, Wu, Zhihao, Jiao, Shuang, Wang, Lijuan, Zou, Yuxia, Jiang, Jingteng, You, Feng
• Format: Journal Article
• Language: English
• Published: Heidelberg Science Press 01.12.2021; Springer Nature B.V; Laboratory for Marine Biology and Biotechnology,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266071,China%CAS and Shandong Province Key Laboratory of Experimental Marine Biology,Center for Ocean Mega-Science,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China; Laboratory for Marine Biology and Biotechnology,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266071,China; University of Chinese Academy of Sciences,Beijing 100049,China%Weihai Fishery Technology Promotion Station,Weihai 264200,China; Shenghang Aquatic Science and Technology Co.,Ltd.,Weihai 264200,China; CAS and Shandong Province Key Laboratory of Experimental Marine Biology,Center for Ocean Mega-Science,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China
• Subjects: Aldehydes; Animal embryos; Aquaculture; CRISPR; Earth and Environmental Science; Earth Sciences; Economics; Editing; Embryos; Fluorescence; Genes; Genetics; Genomes; Green fluorescent protein; In vivo methods and tests; Larvae; Marine fishes; Meteorology; Microinjection; Mutation; Nucleotide sequence; Oceanography; Paralichthys olivaceus; needle with brake; olive flounder; microinjection; CRISPR/Cas9; myomaker; CRISPR /Cas9; gsdf
• ISSN: 1672-5182; 1993-5021; 1672-5174
• DOI: 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.