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<i>Vibrio mimicus</i> is a serious pathogen in aquatic animals, resulting in significant economic losses. The cAMP receptor protein (CRP) often acts as a central regulator in highly pathogenic pathogens. <i>V. mimicus</i> SCCF01 is a highly pathogenic strain isolated from yellow catfish; the <i>crp</i> gene deletion strain (Δ<i>crp</i>) was constructed by natural transformation to determine whether this deletion affects the virulence phenotypes. Their potential molecular connections were revealed by qRT-PCR analysis. Our results showed that the absence of the <i>crp</i> gene resulted in bacterial and colony morphological changes alongside decreases in bacterial growth, hemolytic activity, biofilm formation, enzymatic activity, motility, and cell adhesion. A cell cytotoxicity assay and animal experiments confirmed that <i>crp</i> contributes to <i>V. mimicus</i> pathogenicity, as the LD₅₀ of the Δ<i>crp</i> strain was 73.1-fold lower compared to the WT strain. Moreover, qRT-PCR analysis revealed the inhibition of type II secretion system genes, flagellum genes, adhesion genes, and metalloproteinase genes in the deletion strain. This resulted in the virulence phenotype differences described above. Together, these data demonstrate that the <i>crp</i> gene plays a core regulatory role in <i>V. mimicus</i> virulence and pathogenicity.