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Nucleic Acids Research 2004 32(12):3752-3759; doi:10.1093/nar/gkh715
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Published online 14 July 2004

Nucleic Acids Research, Vol. 32 No. 12 © Oxford University Press 2004; all rights reserved

Inhibition of HIV-1 multiplication by antisense U7 snRNAs and siRNAs targeting cyclophilin A

Songkai Liu, Maria Asparuhova, Vincent Brondani1, Ingrid Ziekau1, Thomas Klimkait1 and Daniel Schümperli*

Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012 Bern, Switzerland and 1 Institute of Medical Microbiology, University of Basel, Petersplatz 10, 4003 Basel, Switzerland

* To whom correspondence should be addressed. Tel: +41 31 631 4675; Fax: +41 31 631 4616; Email: daniel.schuemperli{at}izb.unibe.ch

Received as resubmission July 1, 2004; Accepted July 2, 2004

Human immunodeficiency virus 1 (HIV-1) multiplication depends on a cellular protein, cyclophilin A (CyPA), that gets integrated into viral particles. Because CyPA is not required for cell viability, we attempted to block its synthesis in order to inhibit HIV-1 replication. For this purpose, we used antisense U7 small nuclear RNAs (snRNAs) that disturb CyPA pre-mRNA splicing and short interfering RNAs (siRNAs) that target CyPA mRNA for degradation. With dual-specificity U7 snRNAs targeting the 3' and 5' splice sites of CyPA exons 3 or 4, we obtained an efficient skipping of these exons and a strong reduction of CyPA protein. Furthermore, short interfering RNAs targeting two segments of the CyPA coding region strongly reduced CyPA mRNA and protein levels. Upon lentiviral vector-mediated transduction, prolonged antisense effects were obtained for both types of antisense RNAs in the human T-cell line CEM-SS. These transduced CEM-SS cells showed a delayed, and for the siRNAs also reduced, HIV-1 multiplication. Since the two types of antisense RNAs function by different mechanisms, combining the two approaches may result in a synergistic effect.


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