Endogenous expression of a high-affinity pseudoknot RNA aptamer suppresses replication of HIV-1.
Aptamers, small oligonucleotides derived from an in vitro evolution process called SELEX, are promising therapeutic and diagnostic agents. Although very effective in vitro, only a few examples are available showing their potential in vivo. We have analyzed the effect of a well characterized pseudoknot RNA aptamer selected for tight binding to human immunodeficiency virus (HIV) type 1 reverse transcriptase on HIV replication. Transient intracellular expression of a chimeric RNA consisting of the human initiator tRNA(Met) (tRNA(Meti))/aptamer sequence in human 293T cells showed inhibition of HIV particle release by >75% when the cells were co-transfected with proviral HIV-1 DNA. Subsequent virus production of human T-lymphoid C8166 cells, infected with viral particles derived from co-transfected 293T cells, was again reduced by >75% as compared with the control. As the observed effects are additive, in this model for virus spread, the total reduction of HIV particle formation by transient intracellular expression of the pseudoknot RNA aptamer amounts to >95%. Low-dose HIV infection of human T cells stably expressing the aptamer did not show any virus replication over a period of 35 days. This is the first example of an RNA aptamer selected against a viral enzyme target to show powerful antiviral activity in HIV-1-permissive human T-lymphoid cell lines.
Chaloin
Laurent
L
Max-Planck-Institut für molekulare Physiologie, Abteilung Physikalische Biochemie, Otto Hahn Strasse 11, 44227 Dortmund, Germany.
Lehmann
Maik Jörg
MJ
Sczakiel
Georg
G
Restle
Tobias
T
D001483:Base Sequence; D001667:Binding, Competitive; D002460:Cell Line; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication / Q000235:genetics*
D009843:Oligoribonucleotides; D012358:RNA, Transfer, Met; D012313:RNA; D012194:RNA-Directed DNA Polymerase