title: In vitro and in vivo cleavage of HIV-1 RNA by new SOFA-HDV ribozymes and their potential to inhibit viral replication.
abstract: RNA-based compounds are promising agents to inactivate viruses. New specific hepatitis delta virus (HDV)-derived ribozymes are natural molecules that can be engineered to specifically target a viral RNA. We have designed specific on-off adaptor (SOFA)-HDV ribozymes targeting the tat and rev sequences of the human immunodeficiency virus type 1 (HIV-1) RNA. We show that the SOFA-HDV ribozymes cleave their RNA target in vitro. They inhibit the Tat-mediated transactivation of HIV-1 from 62% to 86% in different assays. In vivo, the amount of HIV RNA was decreased by 60 and 86% with two distinct ribozymes, which indicates that the inhibition of HIV production is directly correlated to the decline in spliced and unspliced viral RNAs. These SOFAHDV- ribozymes inhibited the expression and the viral production of four HIV-1 strains, indicating an extended potential to act on multiple HIV variants. In HEK 293T and HeLa cells transfected with pNL4-3 and the SOFA-HDV-ribozymes, the reduced RNA levels consequently decreased the Gag protein expression in the cell and virus production in the supernatant. When transfected before HIV-1 infection, the ribozymes prevented the incoming virus from being expressed. The ribozymes inhibited HIV production up to 90% when transfected in combination with the HIV protease inhibitor Atazanavir. Our results strongly suggest that SOFA-HDV ribozymes have a great potential to target HIV-1 and to be used as therapeutic agents in combination therapy.
authors.affiliation: Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, McGill University, Montréal, Canada.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Lainé
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Scarborough
authors.forename: Dominique
authors.initials: D
authors.lastname: Lévesque
authors.forename: Ludovic
authors.initials: L
authors.lastname: Didierlaurent
authors.forename: Kaitlin J
authors.initials: KJ
authors.lastname: Soye
authors.forename: Marylène
authors.initials: M
authors.lastname: Mougel
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Perreault
authors.forename: Anne
authors.initials: A
authors.lastname: Gatignol
mesh_terms: D000069446:Atazanavir Sulfate; D001483:Base Sequence; D015683:Gene Products, gag / Q000096:biosynthesis / Q000235:genetics; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D015658:HIV Infections / Q000235:genetics; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D006367:HeLa Cells; D003698:Hepatitis Delta Virus / Q000201:enzymology / Q000235:genetics; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
chemical_list: D015683:Gene Products, gag; D015694:Gene Products, rev; D009842:Oligopeptides; D011725:Pyridines; D016337:RNA, Catalytic; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus; D000069446:Atazanavir Sulfate