screening platform to identify antiviral compounds

Inhibiting the interaction of the human cytidine-deaminase APOBEC3G (A3G) with the HIV-1 specific viral infectivity factor (Vif) represents a novel therapeutic approach where a cellular factor with potent antiviral activity (A3G) plays a key role. In HIV infected cells the interaction of Vif with A3G leads to the subsequent degradation of A3G by the 26S proteasome via the ubiquitin pathway and to the loss of antiviral activity. To establish a stable and convenient cellular testing platform for a high throughput screening of potential antiviral compound libraries, we engineered a double transgenic cell line constitutively expressing an EYFP-A3G fusion as well as Tet-Off controlable Vif protein. With this cell line we are able to measure precisely the Vif-induced degradation of A3G under the presence of potential antiviral compounds in an easy to handle, robust and practical high throughput multi-well plate format with an excellent screening window coefficient (Z-factor) of 0.67.