The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway.  Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly reveals a target for anti-HIV drug development.  In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA   nuclear export.  This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner.  Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP.  The Z' score of this model is 0.822 0.  Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition.  IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.