OBJECTIVE To detect the effect of shRNA inhibiting the expression of NEF in hele cells.METHODS Four pairs of shRNA were transfect four recombinant plasmids into Hele cells.The controls were established,and the RNA and protein were extracted.The primers of NEF and GAPDH were designed.The gene expression of NEF was detected by qPCR,and the protein expression was detected by western blotting.RESULTS The result of qPCR and western blotting indicated the NEF-shRNA-1,NEF-shRNA-2,NEF-shRNA-3 and NEF-shRNA-4 interfering veetors could inhibit NEFmRNA expression.The inhibition rate ofNEF-shRNA-3 was 87%.The inhibition rate of NEF protein of NEF-shRNA-3 was 87% detected by western blotting.CONCLUSION shRNA could inhibit the replication of HIV.