Specific HIV-1 env gene silencing by small interfering RNAs in human peripheral blood mononuclear cells

• Published in: Gene therapy Vol. 10; no. 24; pp. 2046 - 2050
• Main Authors: PARK, W-S, HAYAFUNE, M, MIYANO-KUROSAKI, N, TAKAKU, H
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.11.2003
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Antisense RNA; Antiviral agents; Applied cell therapy and gene therapy; Base Sequence; Binding sites; Biological and medical sciences; Biotechnology; Care and treatment; CD4 antigen; Cells, Cultured; COS Cells; Double-stranded RNA; Env gene; Env protein; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Silencing; Gene therapy; Genes, env - genetics; Genetic aspects; Genetic Therapy - methods; Glycoprotein gp120; Health aspects; Health. Pharmaceutical industry; HIV; HIV infection; HIV-1 - genetics; HIV-1 - physiology; Human immunodeficiency virus; Humans; Industrial applications and implications. Economical aspects; Kinases; Leukocytes (mononuclear); Leukocytes, Mononuclear - virology; Mammalian cells; Medical sciences; Methods; Molecular Sequence Data; Peripheral blood mononuclear cells; Physiological aspects; RNA Interference; RNA, Messenger - genetics; RNA, Small Interfering - genetics; RNA, Viral - genetics; RNA-mediated interference; siRNA; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Virus Replication; Japan; Human; small interfering RNAs; RNA interference; RNA; antiviral RNAi technology; PBMCs; Gene silencing; Blood cell; Mononuclear cell; Gene; Technology; Antiviral; HIV-1 env genes; Gene therapy
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302099

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.