Abstract:
In this study, the suppressor of cytokine signaling 1 (SOCS1) genes
socs1
a and
socs1
b of
Megalobrama amblycephala were used as genetic editing objects, and we screened out appropriate target synthetic guide RNA (gRNA) by online analysis. According to the mixture injection (gRNA and Cas9 protein) in the 1-2 cell stage embryos, the results of identifications of
socs1 expression level by qRT-PCR and gene mutants by sequencing showed that we successfully established
socs1 knock-out mutant. Compared with wild type, the growth performance and body mass of SOCS1a
+/− and SOCS1b
+/− were significantly increased, Meanwhile, the expression levels of inflammatory cytokines
TNF-α and
IL-6 were significantly increased, while the expression levels of
IL-1
β were not changed. After
Aeromonas hydrophila injection, in contrast to the wild type, significant increases in levels of
IL-6 and
TNF-α mRNA were observed in both
socs1
a and
socs1
b heterozygous mutants. The duplicated
socs1 knockout blunt snout bream has been successfully obtained by the CRISPR/Cas9 gene editing system, which provides a basis for further study of the
socs1 gene. Meanwhile, our experimental results will provide a basis and reference for the CRISPR/Cas9 gene editing techniques in other aquaculture species.