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Molecular Medicine

, Volume 18, Issue 1, pp 111–122 | Cite as

Inhibition of Human Immunodeficiency Virus Replication by Cell Membrane-Crossing Oligomers

  • Wilfried Posch
  • Stefan Piper
  • Thomas Lindhorst
  • Birgit Werner
  • Adam Fletcher
  • Holger Bock
  • Cornelia Lass-Flörl
  • Heribert Stoiber
  • Doris Wilflingseder
Research Article

Abstract

Although rapidly becoming a valuable tool for gene silencing, regulation or editing in vitro, the direct transfer of small interfering ribonucleic acids (siRNAs) into cells is still an unsolved problem for in vivo applications. For the first time, we show that specific modifications of antisense oligomers allow autonomous passage into cell lines and primary cells without further adjuvant or coupling to a cell-penetrating peptide. For this reason, we termed the specifically modified oligonucleotides “cell membrane-crossing oligomers” (CMCOs). CMCOs targeted to various conserved regions of human immunodeficiency virus (HIV)-1 were tested and compared with nontargeting CMCOs. Analyses of uninfected and infected cells incubated with labeled CMCOs revealed that the compounds were enriched in infected cells and some of the tested CMCOs exhibited a potent antiviral effect. Finally, the CMCOs did not exert any cytotoxicity and did not inhibit proliferation of the cells. In vitro, our CMCOs are promising candidates as biologically active anti-HIV reagents for future in vivo applications.

Notes

Acknowledgments

The authors were supported by the Austrian Science Found (FWF; project P22165 to D Wilflingseder), the Tyrolean Science Fund (TWF; project D-155140-016-011 to W Posch) and the Bridge Project (Austrian Research Promotion Agency [FFG]; project 815463 to H Stoiber). We would like to thank Christoph Gassner from the local blood bank in Innsbruck, Austria, and CIT Safety and Health Research Laboratories, France, for performing the CMCO stability tests. The reagent ARP033 (M8166 cells) was obtained from the Programme EVA Center for AIDS Reagents (NIBSC, UK) supported by the EC FP6/7 Europrise Network of Excellence, AIDS Vaccine Integrated Project (AVIP) and “Next Generation HIV-1 Immunogens inducing broadly reactive Neutralising antibodies” (NGIN) consortia and the Bill and Melinda Gates GHRC-CAVD Project and was donated by P Clapham. The CMCOs used in this study were already patented (Lindhorst et al. [18], WO 2008/009470; see http://www.wipo.int/patentscope/search/en/WO2008009470). T Lindhorst and H Bock are owners in ugichem GmbH.

Supplementary material

10020_2012_1801111_MOESM1_ESM.pdf (1.4 mb)
Inhibition of Human Immunodeficiency Virus Replication by Cell Membrane-Crossing Oligomers

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Authors and Affiliations

  • Wilfried Posch
    • 1
    • 2
  • Stefan Piper
    • 3
  • Thomas Lindhorst
    • 3
  • Birgit Werner
    • 3
  • Adam Fletcher
    • 2
  • Holger Bock
    • 3
  • Cornelia Lass-Flörl
    • 1
  • Heribert Stoiber
    • 4
  • Doris Wilflingseder
    • 1
    • 2
  1. 1.Department of Hygiene, Microbiology and Social Medicine, Division of Hygiene and Medical MicrobiologyInnsbruck Medical UniversityInnsbruckAustria
  2. 2.MRC/UCL Centre for Medical Molecular Virology, Division of Infection and ImmunityUniversity College LondonLondonUK
  3. 3.Ugichem GmbHInnsbruckAustria
  4. 4.Department of Hygiene, Microbiology and Social Medicine, Division of VirologyInnsbruck Medical UniversityInnsbruckAustria

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