Volume 6 Supplement 2

Frontiers of Retrovirology: Complex retroviruses, retroelements and their hosts

Open Access

Inhibition of HIV-1 expression and replication by SOFA-HDV ribozymes against Tat and Rev mRNA sequences

  • Sébastien Lainé1, 2, 4,
  • Robert J Scarborough1, 2,
  • Dominique Lévesque5,
  • Ludovic Didierlaurent6,
  • Kaitlin J Soye1, 2,
  • Marylène Mougel6,
  • Jean-Pierre Perreault5 and
  • Anne Gatignol1, 2, 3
Retrovirology20096(Suppl 2):P47

https://doi.org/10.1186/1742-4690-6-S2-P47

Published: 24 September 2009

Background

RNA-based compounds are promising methods 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 adapted (SOFA) HDV-ribozymes targeting the regions of the HIV-1 RNA in the Tat and Rev sequences.

Results

We show that these SOFA-HDV ribozymes cleave their Tat RNA target in vitro. They inhibit the Tat-mediated transactivation of HIV-1 long terminal repeat by up to 62 and 86% in luciferase and beta-galactosidase assays, respectively. Inactivation of transfected HIV pNL4-3 molecular clone reached a fourfold inhibition by reverse transcriptase assay of the supernatant and an almost undetectable Gag protein synthesis. In vivo RNA cleavage reached 66 and 86% for two of the tested ribozymes showing that the decrease in HIV production is due to the direct decline in spliced and unspliced viral RNA. These SOFA-HDV-ribozymes were able to target four HIV-1 strains, showing an extended potential to act on multiple HIV variants. When transfected before HIV-1 infection, they prevented incoming virus to be expressed.

Conclusion

Our results show that SOFA-HDV-ribozymes show a great potential to target HIV and to be used as therapeutic agents in gene therapy.

Authors’ Affiliations

(1)
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research, McGill University
(2)
Department of Microbiology and Immunology, McGill University
(3)
Experimental Medicine, McGill University
(4)
CNRS UMR 5236, Université de Bordeaux 2
(5)
RNA Group/Groupe ARN, Département de Biochimie, Université de Sherbrooke
(6)
CNRS UMR 5236-UMI/UMII, CPBS - Equipe ''Assemblage et Réplication des Rétrovirus''

Copyright

© Lainé et al; licensee BioMed Central Ltd. 2009

This article is published under license to BioMed Central Ltd.

Advertisement