Specific HIV-1 env gene silencing by small interfering RNAs in human peripheral blood mononuclear cells.
RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA) in the cell, and results in the silencing of homologous gene expression by the specific degradation of an mRNA containing the same sequence. dsRNA-mediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression. Synthetic 21-23 nucleotide (nt) small interfering RNAs (siRNAs) with 2-nt 3' overhangs were recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we show that synthetic siRNAs targeted against the viral structural Env proteins encoded by HIV-1 can specifically suppress the expression of HIV-1 genes. The siRNA-mediated RNAi also had advantages over antisense RNA-mediated inhibition, in terms of both the ease of designing effective antiviral agents and their potency. Especially, our best env-specific siRNAs, E7145 targeted to the central region of the V3 loop and E7490 targeted to the CD4 binding site of conserved regions on gp120, significantly inhibited the HIV-1 gene expression. Furthermore, E7145 and E7490 were effective against HIV-1(NL4-3) replication in PBMCs for a relatively long time (14 days). Therefore, the use of synthetic siRNAs provides a simple, rapid, and cost-effective tool for new anti-HIV-1 gene therapeutics.
Park
W-S
WS
Department of Industrial Chemistry, Faculty of Engineering, Chiba Institute of Technology, Tsudanuma, Narashino, Chiba, Japan.
Hayafune
M
M
Miyano-Kurosaki
N
N
Takaku
H
H
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002478:Cells, Cultured; D020868:Gene Silencing; D015752:Genes, env / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008969:Molecular Sequence Data; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D014779:Virus Replication
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral