Abstract: Objective · To amplify the interferon regulator factor 3 (IRF3) short hairpin RNA (shRNA) virus and investigate the effect of the virus on the nuclear of Irak1bp1 protein in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. Methods · Adenovirus was amplified in HEK293Tcells and the virus titer was detectedTCID 50 assay. The Raw 264.7 cells were randomly divided into four groups including adenovirus (-) LPS (-) group, adenovirus (-) LPS (+) group, adenovirus (+) LPS (-) group and adenovirus (+) LPS (+) group. The of intracellular IRF3 mRNA was detectedreal-time PCR, and the nuclear of IRF3 and Irak1bp1 protein were detectedWestern blotting. Results · The titer of adenovirus was 2.2×1011 PFU/mL and the best MOI was 300. The of IRF3 mRNA and nuclear IRF3 protein in LPS-stimulated Raw 264.7 cells were significantly higher than those of the control group. The cellular constitutive of IRF3 at mRNA level and the LPS-induced ofIRF3 were significantly inhibited after transfection of Raw 264.7 cells with adenovirus strains carrying IRF3 shRNA. However, the nuclear constitutive of IRF3 protein was not affectedIRF3 shRNA in the unstimulated state. The of nuclear Irak1bp1 protein was significantly higher than that of the control group. The nuclear constitutive and the LPS-induced of Irak1bp1 protein were not affectedIRF3 shRNA. Conclusion · Transfection of LPS-stimulated Raw 264.7 cells with adenovirus strains carrying IRF3 shRNA could effectively inhibit the ofIRF3, but not affect the nuclear of Irak1bp1 protein.

Key words: interferon regulator factor 3, short hairpin RNA , interleukin-1 receptor associated kinase 1 binding protein 1 (Irak1bp1), lipopolysaccharide (LPS)