CRISPR/Cas9-mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda

The development of type II clustered regularly interspaced short palindromic repeats (CRISPR) system has resulted in the revolution of genetic engineering and this technology has been applied in genome editing of various species. However, there is no report about the target-specific genome editing in shrimp. In this research, we developed the microinjection method for the ridgetail white prawn Exopalaemon carinicauda and successfully applied the CRISPR/Cas9 technology on the genome editing of E. carinicauda. Through co-injection of mRNA of Cas9 nuclease and gRNA specialized for E. carinicauda chitinase 4 (EcChi4), the shrimp with the indels mutations were obtained. Further analysis showed that the mutations could be transmitted to the next generation. It is the first time that the site-specific genome editing was successfully demonstrated in decapod, which will further contribute to the study of functional genomic in decapods.

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