The Research On Small Interfering RNA Targeting HIV-1Vif Gene

Acquired immunodeficiency syndrome (AIDS) has spread at an alarming ratein the world and caused great damages to human society. since it was first reportedin the early1980s, in the United States. The prevention and treatment has become anurgent social problem. Although the widely used the highly active antiretroviraltherapy (HAART) could improve the life quality of HIV-1infected patientdramatically, treatment was interrupted due to the high cost, drug resistance and sideeffects. To achive sustained antiretrovial effects, new antiretroviral drugs andapproaches are demanded. With the development of molecular biology, stem cell,nanotechnology and other related technologies, gene therapy for HIV-1infection hasattracted considerable attention.HIV-1viral infectivity factor (Vif) plays an important role in the HIV-1infection and disease progression. In this study, RNAi technology was applied tointerfere the expression of HIV-1vif gene to inhibit HIV-1infection. Seven shRNA-expression vectors targeting distinct sites of vif gene sequence were constructed andcotransfected into the293FT cells with vif expression vector. It was shown thatshRNA7and shRNA10could inhibit the expression of HIV-1vif gene by WesternBlot and Real-time PCR. Vectors expressing shRNA7or shRNA10, APOBEC3G,HIV-1bockbone and envelope were cotransfected into the293FT cells to produceHIV-1pseudovirus. The results from p24quatification showed that shRNA had noeffect on the virus packaging. Studies using HIV-1pseudovirus system revealed thatshRNA7could inhibit the infectivity of HIV-1. The inhibitive rate was52.93%and89.45%respectively, with the transfecting dose of0.5g and1.5g. The anti-HIV-1activity was followed by shRNA10. Its inhibitive rate was48.95%, with the dose of1.5g.