title: Conserved sequences in the current strains of HIV-1 subtype A in Russia are effectively targeted by artificial RNAi in vitro.
abstract: Highly active antiretroviral therapy has greatly reduced the morbidity and mortality of AIDS. However, many of the antiretroviral drugs are toxic with long-term use, and all currently used anti-HIV agents generate drug-resistant mutants. Therefore, there is a great need for new approaches to AIDS therapy. RNAi is a powerful means of inhibiting HIV-1 production in human cells. We propose to use RNAi for gene therapy of HIV/AIDS. Previously we identified a number of new biologically active siRNAs targeting several moderately conserved regions in HIV-1 transcripts. Here we analyze the heterogeneity of nucleotide sequences in three RNAi targets in sequences encoding the reverse transcriptase and integrase domains of current isolates of HIV-1 subtype A in Russia. These data were used to generate genetic constructs expressing short hairpin RNAs 28-30-bp in length that could be processed in cells into siRNAs. After transfection of the constructs we observed siRNAs that efficiently attacked the selected targets. We expect that targeting several viral genes important for HIV-1 reproduction will help overcome the problem of viral adaptation and will prevent the appearance of RNAi escape mutants in current virus strains, an important feature of gene therapy of HIV/AIDS.
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia. Electronic address: tchurikov@eimb.ru.
authors.forename: Nickolai A
authors.initials: NA
authors.lastname: Tchurikov
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Daria M
authors.initials: DM
authors.lastname: Fedoseeva
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Natalya M
authors.initials: NM
authors.lastname: Gashnikova
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Sosin
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Gorbacheva
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Ildar R
authors.initials: IR
authors.lastname: Alembekov
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Vladimir R
authors.initials: VR
authors.lastname: Chechetkin
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Yuri V
authors.initials: YV
authors.lastname: Kravatsky
authors.affiliation: Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Kretova
mesh_terms: D001483:Base Sequence; D017124:Conserved Sequence; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012426:Russia; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering
keywords: Genetic constructs; HIV-1 in Russia; RNAi; RNase protection; Subtype A; Transfection; siRNAs