title: A Conserved Target Site in HIV-1 Gag RNA is Accessible to Inhibition by Both an HDV Ribozyme and a Short Hairpin RNA.
abstract: Antisense-based molecules targeting HIV-1 RNA have the potential to be used as part of gene or drug therapy to treat HIV-1 infection. In this study, HIV-1 RNA was screened to identify more conserved and accessible target sites for ribozymes based on the hepatitis delta virus motif. Using a quantitative screen for effects on HIV-1 production, we identified a ribozyme targeting a highly conserved site in the Gag coding sequence with improved inhibitory potential compared to our previously described candidates targeting the overlapping Tat/Rev coding sequence. We also demonstrate that this target site is highly accessible to short hairpin directed RNA interference, suggesting that it may be available for the binding of antisense RNAs with different modes of action. We provide evidence that this target site is structurally conserved in diverse viral strains and that it is sufficiently different from the human transcriptome to limit off-target effects from antisense therapies. We also show that the modified hepatitis delta virus ribozyme is more sensitive to a mismatch in its target site compared to the short hairpin RNA. Overall, our results validate the potential of a new target site in HIV-1 RNA to be used for the development of antisense therapies.
authors: 1] Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, Montréal, Québec, Canada [2] Department of Microbiology & Immunology, McGill University, Montréal, Québec, Canada.
authors: Robert J
authors: RJ
authors: Scarborough
authors: Département de Biochimie, RNA Group/Groupe ARN, Université de Sherbrooke, Sherbrooke, Québec, Canada.
authors: Michel V
authors: MV
authors: Lévesque
authors: 1] Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, Montréal, Québec, Canada [2] Department of Microbiology & Immunology, McGill University, Montréal, Québec, Canada.
authors: Etienne
authors: E
authors: Boudrias-Dalle
authors: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors: Ian C
authors: IC
authors: Chute
authors: 1] Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, Montréal, Québec, Canada [2] Department of Microbiology & Immunology, McGill University, Montréal, Québec, Canada.
authors: Sylvanne M
authors: SM
authors: Daniels
authors: Atlantic Cancer Research Institute, Moncton, New Brunswick, Canada.
authors: Rodney J
authors: RJ
authors: Ouellette
authors: Département de Biochimie, RNA Group/Groupe ARN, Université de Sherbrooke, Sherbrooke, Québec, Canada.
authors: Jean-Pierre
authors: JP
authors: Perreault
authors: 1] Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, Montréal, Québec, Canada [2] Department of Microbiology & Immunology, McGill University, Montréal, Québec, Canada [3] Department of Medicine, McGill University, Montréal, Québec, Canada.
authors: Anne
authors: A
authors: Gatignol