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  • Oral presentation
  • Open Access

Towards gene therapy against HIV-1: new therapeutic target in gag RNA accessible to ribozymes and RNA interference molecules

  • 1,
  • 1,
  • 2,
  • 1,
  • 3,
  • 1,
  • 3 and
  • 2
BMC Infectious Diseases201414 (Suppl 2) :O19

https://doi.org/10.1186/1471-2334-14-S2-O19

  • Published:

Keywords

  • Combination Gene
  • Hepatitis Delta Virus
  • Lymphocytic Cell
  • Transfected HEK293T Cell
  • Antisense Molecule

Aim

Antisense molecules targeting HIV-1 RNA have the potential to be used as part of combination gene or drug therapy to treat HIV-1 infection to reach a functional or complete cure. Only a small number of extremely active molecules currently exist and a treatment option has not yet been identified in clinical trials. We have previously developed new hepatitis delta virus (HDV)-derived ribozymes (Rzs) called "switch on-off adaptor" (SOFA) to target HIV-1 RNA [1]. Our aim is to develop highly active RNA-based molecules with complementary mechanisms, which are able to reach a large number of HIV-1 variants.

Methods

We screened HIV-1 RNA to identify conserved target sites for new HDV-Rzs [2]. We designed new SOFA-HDV-Rzs against the Gag RNA and developed a rapid test to evaluate the inhibition of HIV-1 production. We designed small interfering (si) RNAs targeting the same region and tested their activity on HIV-1 replication.

Results

We identified 13 conserved regions in the gag RNA and constructed the corresponding Rzs. We transfected HEK293T cells with these Rzs and HIV-1 molecular clones. We identified one Rz that was particularly efficacious. We then constructed siRNAs and short hairpin (sh) RNAs targeting the same sequence. The shRNA was very active against HIV-1 clades B, C and A/G. Neither the Rz, nor the shRNA disturbs the cellular transcriptome, suggesting no toxicity. In lymphocytic cell lines, both the Rz and the shRNA inhibit long-term HIV-1 replication [2].

Conclusions

We identified new SOFA-HDV-Rz, siRNA and shRNA targeting HIV-1 Gag RNA. The shRNA is as active as the only shRNA that has advanced to clinical trials and targets more strains. Long-term inhibitory activity of these molecules shows that this site is particularly accessible to other antisense molecules. These molecules have a high potential to be used in combination gene therapy or as drugs with appropriate delivery methods.

Authors’ Affiliations

(1)
Lady Davis Institute for Medical Research, McGill University, Montreal, Canada
(2)
Department of Biochemistry, University of Sherbrooke, Sherbrooke, Canada
(3)
Atlantic Institute for research on cancer, Moncton, Canada

References

  1. Lainé S, et al: RNA Biol. 2011, 8: 343-Google Scholar
  2. Scarborough RJ, et al: Methods Mol Biol. 2014, 1103: 31-Google Scholar

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