Aptamers directed to HIV-1 reverse transcriptase display greater efficacy over small hairpin RNAs targeted to viral RNA in blocking HIV-1 replication.
RNA molecules can be powerful inhibitors of HIV-1 replication. To determine the relative efficacy of siRNAs and RNA aptamers, a direct comparison of three anti-HIV reverse transcriptase aptamers and three shRNAs targeted to HIV-1(R3b) was made. U6 promoter-driven anti-HIV genes were delivered into CEMx174 cells via a retroviral vector, and transduced cells were sorted out via green fluorescent protein function and challenged with HIV. The results show that, at low virus input, shRNAs can block HIV as efficiently as aptamers. When expressed in target cells, both classes of inhibitors blocked early events of reverse transcription, suggesting they are both able to access intracellular reverse transcription complexes. However, at higher multiplicities of infection (m.o.i. of 50), while the aptamers could efficiently inhibit HIV replication, shRNAs did not. RNase protection assays indicated similar steady-state levels or nucleocytoplasmic distribution showing that the differential efficacy was not a reflection of intracellular concentration. The higher potency of anti-RT aptamers could be due to their ability to inhibit two successive rounds of reverse transcription owing to their unique ability to be encapsidated into virion particles. Furthermore, anti-RT aptamers expressed in T cells afforded protection against high-dose infection by chimeric RT-SHIV viruses.
Joshi
Pheroze J
PJ
Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, GB401, Bronx, NY 10461, USA.
North
Thomas W
TW
Prasad
Vinayaka R
VR
D020029:Base Pairing; D002460:Cell Line; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D007700:Kinetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000235:genetics; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics; D014779:Virus Replication
D012342:RNA, Small Nuclear; D012367:RNA, Viral; C405486:U6 small nuclear RNA; D012313:RNA; D054303:HIV Reverse Transcriptase