Expression of Chimeric Neo-Rev Response Element Sequences Interferes with Rev-Dependent HIV-1 Gag Expression

Recombinant plasmids containing reiterated human immunodeficiency virus type 1 (HIV-1) Rev response element (RRE) sequences were constructed to suppress Rev-dependent HIV-1 Gag expression. The mammalian expression vectors pMAMneo containing one, three, or six repeats of the RRE sequence were cotransfected with a HIV-1 HTLV-III_B proviral DNA into HeLa cells. All three RRE expression plasmids reduced replication of HIV-1 with similar efficacy. Furthermore, the chimeric expression vector pCMVneoRRE6×→(containing six copies of the RRE sequence) was used to etablish HeLa cell lines constitutively expressing RRE. A plasmid encoding a Rev-dependent HIV-1 p24 Gag protein was cotransfected with the wild-type Rev expression plasmid into three different RRE-expressing HeLa cell lines. p24 Gag protein production in the culture supernatants of the HeLaneoRRE cells was compared with two neo-expressing cell lines. Although all cell lines (HeLaneoRRE, HeLaneo) displayed similar transfection efficiencies, p24 Gag protein synthesis was markedly reduced in the RRE-expressing cell lines in comparison to the control cells.

Overview summary

Human immunodeficiency virus type 1 (HIV-1) is the causative agent of acquired immunodeficiency syndrome (AIDS) but currently no adequate treatment for this disease is available. Intracellular immunization is a novel approach to achieve inhibition of HIV-1 replication. Several investigators described different strategies employing trans-dominant mutants of HIV-1 gene products, antisense RNA, ribozymes, and overexpression of TAR sequences as decoys to inhibit HIV-1 replication. Zimmermann et al. demonstrate that the use of Rev response element (RRE) sequences to interfere with the HIV-1 Rev trans-activator protein activity represents an alternative approach for anti-retroviral gene intervention.