Identification of aptamers against the DNA template for in vitro transcription of the HIV-1 TAR element.

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Identification of aptamers against the DNA template for in vitro transcription of the HIV-1 TAR element.

Antisense Nucleic Acid Drug Dev. 1997 Aug;7(4):369-80. Unique Identifier : AIDSLINE MED/97448864
Boiziau C; Dausse E; Mishra R; Duconge F; Toulme JJ; INSERM U 386, IFR Pathologies Infectieuses, Universite Victor Segalen,; Bordeaux, France.


Abstract: We have extracted from a random population of about 10(9) oligodeoxynucleotides a series of 21-mers that are able to bind to a folded DNA 76-mer used as a template for in vitro transcription of the TAR element of the retrovirus HIV-1, by the T7 RNA polymerase. Five aptastrucs, that is, aptamers able to bind to the structure, out of 15 analyzed sequences, share the consensus motif 5'-PyGGG(TG)PyC, complementary in part to a weak double-stranded region of the target. (The parentheses indicate that either T or G is missing in one of these aptastrucs.) A dissociation constant of about 3 microM was evaluated by electrophoretic mobility shift assay for the winner sequence. Interactions between the aptastruc and the target sequences involve more than Watson-Crick base pairing of the consensus octamer. The binding is chemistry dependent. Phosphorothioate oligodeoxyribonucleotides and 2'-O-methyl oligoribonucleotides derived from the selected aptastrucs exhibit a weak if any affinity for the target.
Keywords: *HIV Long Terminal Repeat *HIV-1/GENETICS *Oligonucleotides, Antisense/PHARMACOLOGY *Transcription, Genetic/DRUG EFFECTSKWDhivlongterminalrepeatKWDhiv-1/geneticsKWDoligonucleotides,antisense/pharmacologyKWDtranscription,genetic/drugeffects
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