Specific inhibition of in vitro reverse transcription using antisense oligonucleotides targeted to the TAR regions of HIV-1 and HIV-2.
Antisense oligonucleotides (ODNs) overlapping the stem-loop structure of the trans-activating responsive (TAR) element at the 5' end of HIV-1 and HIV-2 viral RNAs were tested for their inhibitory effect on cDNA synthesis by HIV-1 and HIV-2 reverse transcriptases (RT). Inhibition of reverse transcription is sequence-specific and enhanced by the presence of the RT-associated RNase H activity. The degree of inhibition obtained with the anti-TAR antisense is significantly higher than with other HIV-1 targeted antisense ODNs used before [1]. Gel retardation showed a stable specific complex between the 16- and 25-mer anti-TAR HIV-1 selected ODNs and the target region. No complex was observed with a non-inhibitor 22-mer anti-TAR ODN and with the corresponding control sequences. Targeting of the first stem-loop in the 5' region of HIV-2 RNA by anti-TAR ODNs inhibited very strongly reverse transcription by HIV-2 RT. The structure of the antisense and the target sequence affect annealing efficiency and hence the degree of inhibition of reverse transcription.
Boulme
F
F
Laboratoire Reger, Institut de Biochimie et Génétique Cellulaires du CNRS, IBGC-CNRS, Bordeaux, France.
Perälä-Heape
M
M
Sarih-Cottin
L
L
Litvak
S
S
D001483:Base Sequence; D018076:DNA, Complementary / Q000096:biosynthesis / Q000187:drug effects; D004279:DNA, Viral / Q000096:biosynthesis / Q000187:drug effects; D004586:Electrophoresis / Q000379:methods; D016325:HIV Long Terminal Repeat / Q000187:drug effects* / Q000235:genetics; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology*; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000187:drug effects / Q000235:genetics*; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D016914:Ribonuclease H / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
D018076:DNA, Complementary; D004279:DNA, Viral; D009692:Nucleic Acid Heteroduplexes; D016376:Oligonucleotides, Antisense; D012367:RNA, Viral; D018894:Reverse Transcriptase Inhibitors; C072416:reverse transcriptase, Human immunodeficiency virus 2; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D016914:Ribonuclease H