Objective:To construct interferon regulatory factor-1(IRF-1) and its shRNA expression vectors,and to assess their function in rat glomerular mesangial cell (GMC). Methods:The full-length IRF-1 or three 19~21 bp reverse repeated motifs targeting of IRF-1 gene were synthesized and cloned into eukaryotic expression plasmid pcDNA3.1/myc and pGCsi.U6.neo.GFP. After screened and confirmed,the recombinant plasmids were transfected into GMC,then Western blot was used to detect the expression of IRF-1 in GMC and find out the optimal shRNA. Results:It was verified by partial nucleotide sequencing and restriction endonuclease digestion that the constructed eukaryotic vectors were correct. The results by Western blot showed that the constructed pcDNA3.1/IRF-1 plasmid expressed correctly and the IRF-1 shRNA-2 was optimal shRNA,which effectively silenced the target gene. Conclusion:The pcDNA3.1/IRF-1 plasmid and its shRNA were constructed successfully. These data provide the essential experimental tools for studying biological functions of IRF-1 gene in the future.