Endogenous expression of an anti-TAR aptamer reduces HIV-1 replication.
An anti-TAR RNA aptamer called R06, which binds tightly and specifically to the trans-activation responsive (TAR) element of the human immunodeficiency virus type 1 (HIV-1) through loop-loop interactions has been previously selected.(1) We used HIV-based retroviral vectors to express the R06 aptamer. Its synthesis was driven by the U16 snoRNA. We investigated the ability of this cassette to interfere with TAR-mediated transcription using HeLa P4 cells stably expressing the beta-galactosidase gene under the control of the HIV-1 5'LTR. We demonstrated that, upon HIV-1 infection, the beta-galactosidase activity was reduced in cells expressing the nucleolar U16-R06 transcript. The replication of HIV-1 in these cells was also reduced as shown by quantification of the HIV-1 protease gene 24 h post-infection. This effect was specific and related to the formation of R06 TAR complex as an aptamer with a mutated loop, which was no longer able to bind to TAR, did not show any effect. The nucleolus is likely a compartment of interest for targeting the TAR-protein complex responsible for the trans-activation of transcription of the HIV-1 genome.
Kolb
Gaëlle
G
INSERM U 869, Institut Européen de Chimie et Biologie, Pessac, France.
Reigadas
Sandrine
S
Castanotto
Daniela
D
Faure
Aurélie
A
Ventura
Michel
M
Rossi
John J
JJ
Toulmé
Jean-Jacques
JJ
D019380:Anti-HIV Agents / Q000737:chemistry* / Q000378:metabolism*; D052157:Aptamers, Nucleotide / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D015533:Transcriptional Activation / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
D019380:Anti-HIV Agents; D052157:Aptamers, Nucleotide