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Nucleic Acids Research, Vol 26, Issue 10 2273-2278, Copyright © 1998 by Oxford University Press

ARTICLES

Selective inhibition of cell-free translation by oligonucleotides targeted to a mRNA hairpin structure

R Le Tinevez, RK Mishra and JJ Toulme
INSERM U 386, IFR Pathologies Infectieuses, Universite Victor Segalen, 146 rue Leo Saignat, 33076 Bordeaux cedex, France.

Using an in vitro selection approach we have previously isolated oligodeoxy aptamers that can bind to a DNA hairpin structure without disrupting the double-stranded stem. We report here that these oligomers can bind to the RNA version of this hairpin, mostly through pairing with a designed 6 nt anchor. The part of the aptamer selected against the DNA hairpin did not increase stability of the RNA-aptamer complex. However, it contributed to the binding site for Escherichia coli RNase H, leading to very efficient cleavage of the target RNA. In addition, a 2'- O -methyloligoribonucleotide analogue of one selected sequence selectively blocked in vitro translation of luciferase in wheat germ extract by binding to the hairpin region inserted upstream of the initiation codon of the reporter gene. Therefore, non- complementary oligomers can exhibit antisense properties following hybridization with the target RNA. Our study also suggests that in vitro selection might provide a means to extend the repertoire of sequences that can be targetted by antisense oligonucleotides to structured RNA motifs of biological importance.
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