期刊
FISH & SHELLFISH IMMUNOLOGY
卷 97, 期 -, 页码 421-431出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fsi.2019.12.037
关键词
Exopalaemon carinicauda; Carotenoid isomerooxygenase; CRISPR/Cas9
资金
- National Key R&D Program of China [2018YFD0900205]
- National Natural Science Foundation of China [31872613, 41876196]
- Natural Science Foundation of Hebei Province of China [C2019201236, D2019201239]
- Innovation & entrepreneurship training program for college students of Hebei Province [S201910075061]
- Innovation & entrepreneurship training program for college students of Hebei University [2019177]
- Hangzhou Qianjiang Special Expert for Jiquan Zhang
During the immune defense reaction of invertebrate, a plenty of reactive oxygen species (ROS) could be induced to product. Though ROS can kill foreign invaders, the accumulation of these reactive molecules in animals will cause serious cell damage. Carotenoids could function as scavengers of oxygen radicals. In this research, cDNA and genomic DNA of one carotenoid isomerooxygenase gene (named EcNinaB-X1) were cloned from Exopalaemon carinicauda. EcNinaB-X1 gene was composed of 12 exons and 11 introns. EcNinaB-X1 knock-out (KO) prawns were produced via CRISPR/Cas9 technology and the change of their phenotypes were analyzed. Of the 400 injected one-cell stage embryos with cas9 mRNA and one sgRNA targeting the first exon of EcNinaB-X1 gene, 26 EcNinaB-X1-KO prawns were generated and the mutant rate reached 6.5% after embryo injection. The EcNinaB-X1-KO prawns had significant lower mortality than those in wild-type group when the prawns were challenged with Vibrio parahaemolyticus or Aeromonas hydrophila. In conclusion, we first demonstrate the function of the carotenoid isomerooxygenase gene in immune defense of E. carinicauda by performing directed, heritable gene mutagenesis.
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