A quantitative GFP-based bioassay for the detection of HIV-1 Tat transactivation inhibitors

The Tat function of the human immunodeficiency virus (HIV) represents an im portant target for the development of new anti-HIV drugs. A rapid, sensitiv e and simple bioassay was developed for the detection of HIV transactivatio n inhibitors. A reporter plasmid based on the expression of the green fluor escent protein (GFP) under control of the HIV-1 long terminal repeat (LTR) was constructed. This reporter gene can be quantified by simply measuring t he fluorescence irradiated by GFP-producing cells, without the need of extr action procedures or enzymatic assays. Cells, stably expressing HIV-1 Tat p rotein, were transfected with this plasmid and the inhibitory effect of ant i-Tat drugs was assessed by measuring the inhibition of fluorescence. Using this assay system the anti-transactivation activity of several known compo unds was confirmed. This is the first HIV transactivation assay using GFP r eporter gene in microtiter plates. The assay can be used for the detection and quantification of HIV trans activation, and for the high throughput eva luation of anti-trans activation drugs in different cellular backgrounds. ( C) 2001 Elsevier Science B.V. All rights reserved.