TARGET-SPECIFIC ARREST OF MESSENGER-RNA TRANSLATION BY ANTISENSE 2'-O-ALKYLOLIGORIBONUCLEOTIDES

H. E. Johansson, Graham Belsham, B. S. Sproat, M. W. Hentze

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

We describe a novel experimental approach to investigate mRNA translation. Antisense 2'-O-allyl oligoribonucleotides (oligos) efficiently arrest translation of targeted mRNAs in rabbit reticulocyte lysate and wheat germ extract while displaying minimal non-specific effects on translation. Oligo/mRNA-hybrids positioned anywhere within the 5' UTR or the first similar to 20 nucleotides of the open reading frame block cap-dependent translation initiation with high specificity. The thermodynamic stability of hybrids between 2'-O-alkyl oligos and RNA permits translational inhibition with oligos as short as 10 nucleotides. This inhibition is independent of RNase H cleavage or modifications which render the mRNA untranslatable. We show that 2'-O-alkyl oligos can also be employed to interfere with cap-independent internal initiation of translation and to arrest translation elongation. The latter is accomplished by UV-crosslinking of psoralen-tagged 2'-O-methyloligoribonucleotides to the mRNA within the open reading frame. The utility of 2'-O-alkyloligoribonucleotides to arrest translation from defined positions within an mRNA provides new approaches to investigate mRNA translation.
Original languageEnglish
JournalNucleic Acids Research
Volume22
Issue number22
Pages (from-to)4591-4598
ISSN0305-1048
DOIs
Publication statusPublished - 1994
Externally publishedYes

Fingerprint Dive into the research topics of 'TARGET-SPECIFIC ARREST OF MESSENGER-RNA TRANSLATION BY ANTISENSE 2'-O-ALKYLOLIGORIBONUCLEOTIDES'. Together they form a unique fingerprint.

Cite this