Inhibition of human immunodeficiency virus type 1 by packageable, multigenic antisense RNA

Viral-based vectors can provide an efficient delivery mechanism for stable expression of antisense RNA. To enhance and propagate the antiviral effect of antisense RNA, two novel human immunodeficiency virus type 1 (HIV-1)-bas ed vector DNAs, designated as pMAG7 and pMAG19, were constructed which cont ained HIV-1 cis-acting packaging elements and produced multigenic HIV-1 ant isense RNA that could target the entire pal, env, vif, vpu, vpr, rev, and t at and portions of gag and nef. The two DNAs were identical except that pMA G19 had additional gag coding sequences, Cotransfection of pMAG DNA and inf ectious, cloned HIV-1 DNA in 293 cells inhibited virus production (81%-98% reduction in reverse transcriptase activity) of various T cell-tropic and m acrophage-tropic clade B isolates, such as NL4-3, YU-2, and JR-CSF, In addi tion, virion-associated pMAG antisense RNA was detected in residual virus p articles produced by pNL4-3 in the presence of pMAG7 DNA, and the antisense sequences were stably transferred by infection of 174 x CEM cells. The res ults suggest that pMAG DNA may confer broad protection against HIV-1 by red ucing initial virus burden due to antisense RNA and subsequent virus spread by propagation of antisense sequences along with wild-type virus.