A CRISPR/Cas9-mediated mutation in chitinase changes immune response to bacteria inExopalaemon carinicauda


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Abstract

Chitinase, belonging to family 18 glycosyl hydrolase, is a multi-gene family and it has many functions. Generation of loss-of-function mutant targeting an interesting gene is a common way to clarify its function based on reverse genetics. In this study, we first reported the immune defense of a chitinase gene (EcChi4) in Exopalaemon carinicauda using its EcChi4-deletion mutant. EcChi4 was predominantly expressed in hepatopancreas and was upregulated after challenge with Vibrio parahaemolyticus or Aeromonas hydrophila. After knockout EcChi4 gene using CRISPR/Cas9 tool, the prawns in EcChi4-deletion group had significant higher mortality than those in wild-type group when the prawns were challenged with V. parahaemolyticus or A. hydrophila. In conclusion, we first demonstrate the function of a chitinase gene in immune defense of E. carinicauda by performing directed, heritable gene mutagenesis. In the future, CRISPR/Cas9 should be widely applicable as a feasible means for gene editing in E. carinicauda for the study of important biological questions that cannot be easily addressed in other decapods.HighlightsEcChi4 was predominantly expressed in hepatopancreas of E. carinicauda.EcChi4 was upregulated after challenge with V. parahaemolyticus or A. hydrophila.EcChi4-deleted prawn had higher mortality than wild-type after bacterial challenge.We first demonstrate the gene function of prawn by performing gene mutagenesis.

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