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

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...

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Bibliographic Details
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
Description
Summary: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.

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