Objective:To construct the recombinant adenoviruses containing microRNA against rat GR.Methods:We designed and synthesised four pairs of oligonucleotide-encoding microRNA against rat GR and cloned them into pcDNATM 6.2-GW/EmGFPmiR to generate expression vectors.The vectors were transfected into rat embryonic fibroblast to detect the effect of interference for initial screening through Western blot.Then the adenoviral expression constructs were generated through BP reaction and LR recombination reaction.The resulting constructs were transfected into 293A cells to package infective adenoviruses.After several rounds of repeated infection of 293A cells

the adenoviruses were amplified to the appropriate titer.The adenoviruses were infected into Leydig cells.Western blot was performed to analysis for knockdown of GR.Results:Leydig cells were efficiently infected with recombinant adenovirus.The rate of infection was 80%.GR expression in Leydig cells infected with miGR adenoviruses(the adenoviruses containing microRNA against rat GR) was significantly decreased compared with that in the Leydig cells of control group.Conclusion:The recombinant miGR adenovirus can effectively interfere with the GR expression in primary rat Leydig cells.