The NF-κB/Rel transcription factors play essential roles in the induction and regulation of innate immune responses. In the present study, the full-length cDNA of CgRel from the Pacific oyster Crassostrea gigas was of 2,647 bp with an RHD and an IPT domain. The mRNA of CgRel was found to be constitutively expressed in all the tested tissues including gills, hepatopancreas, gonad, adductor muscle, labial palps, mantle, hemocytes, and ganglion. After lipopolysaccharide (LPS) stimulation, the expression level of CgRel mRNA in hemocytes was up-regulated to the first peak at 3 h (3.06-fold compared to the control group, p < 0.001) and second peak at 48 h (1.96-fold, p < 0.05). It increased significantly at 3 h (7.68-fold compared to the control group, p < 0.001), 24 h (3.63-fold, p < 0.05) and 48 h (1.99-fold, p < 0.05) post Vibrio splendidus stimulation, respectively. The protein of CgRel was translocated from cytoplasm into nucleus of oyster hemocytes after LPS stimulation. The mRNA expression levels of interleukin17s (CgIL17s) and big defensin (CgBigDef1) in hemocytes were examined after the expression of CgRel was silenced by RNAi. The transcripts of CgIL17-1 (0.25-fold of the control group, p < 0.01), CgIL17-2 (0.12-fold, p < 0.01), CgIL17-4 (0.33-fold, p < 0.01), CgIL17-6 (0.27-fold, p < 0.05) and CgBigDef1 (0.38-fold, p < 0.01) in CgRel-knockdown oysters decreased significantly at 12 h after LPS stimulation. The results indicated that CgRel played important roles in the immune defense against bacteria by regulating the expression of CgIL17 and CgBigDef1.