Exploration The New Approach For AIDS Therapy By ApplyingRNA Interference To Nef Gene Expression

Acquired immunodeficiency syndrome (AIDS) is a kind of disease infected by human immunodeficiency virus (HIV) The nef gene is unique to primate lentiviruses (human immunodeficiency type 1 [HIV-1], HIV-2, and simian immunodeficiency virus [SIV]) and encodes a myristoylated membrane-associated protein of 25 to 34 kDa. Nef is essential for high-level virus replication and full pathogenic potential. The mechanisms by which Nef acts as a positive factor during infection are unclear but are most likely related to both viral and cellular factors. Nef has been shown to enhance virion infectivity and reverse transcriptase activity. At the cellular level, Nef reduces the level of cell surface receptors. These observations imply that nef has a role in perturbing the cell activation pathway(s), which presumably influences viral replication in the host and possibly cause dysfunction of cells in the immune system.Eukaryotic expression plasmid of nef was constructed and transfected into breast cancer line Hela. Three interference plasmids targeted different position of nef were constructed. Transfected efficiency and expression of nef in Hela cell were detected by observe the green fluorescence shRNA-expressing.plasmid and nef plasmid were co-transfected into Hela cell .difference expression of nef was detedted by RT-PCR and Western Blot.The most effective plasmid of the three was choosed to constructed virusplasmid. Virusplasmid was tansfected into packing cell and prouced virus. CEMx174 cell were infected with virus collected and siv. Expression of nef and gag were detedted by RT-PCR and Western Blot.Overall, our results indicate that nef shRNA transduced into lymphocyte cell line inhibited HIV-1 transcription.Nef via RNAi pathways may allow persistently low pathogenic or latent infection of HIV.