Inhibition of HIV-1 multiplication by antisense U7 snRNAs and siRNAs targeting cyclophilin A.
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.
Liu
Songkai
S
Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012 Bern, Switzerland.
Asparuhova
Maria
M
Brondani
Vincent
V
Ziekau
Ingrid
I
Klimkait
Thomas
T
Schümperli
Daniel
D
D002460:Cell Line; D021984:Cyclophilin A / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D005091:Exons; D005822:Genetic Vectors; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D034622:RNA Interference; D012322:RNA Precursors / Q000378:metabolism; D012326:RNA Splicing; D016372:RNA, Antisense / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D012342:RNA, Small Nuclear / Q000037:antagonists & inhibitors*; D013601:T-Lymphocytes / Q000821:virology; D014779:Virus Replication
D012322:RNA Precursors; D016372:RNA, Antisense; D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; C405487:U7 small nuclear RNA; D021984:Cyclophilin A