2′-O-alkyl oligoribonucleotides as antisense probes

Adolfo M. Iribarren, Brian S. Sproat, Philippe Neuner, Ingrid Sulston, Ursula Ryder, Angus I. Lamond

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

2′-O-Methyl oligoribonucleotides have recently been introduced as antisense probes for studying RNA processing and for affinity purification of RNA-protein complexes. To identify RNA analogues with improved properties for antisense analysis, 2′-O-alkyl oligoribonucleotides were synthesized in which the alkyl moiety was either the three-carbon linear allyl group or the five-carbon branched 3,3-dimethylallyl group. Both these analogues were found to be completely resistant to degradation by either DNA- or RNA-specific nucleuses. Use of biotinylated derivatives of the probes to affinity-select ribonucleoprotein particles from crude HeLa cell nuclear extracts showed that the presence of the bulky 3,3-dimethylallyl group significantly reduces affinity selection, whereas the allyl derivative binds rapidly and stably to targeted sequences and affinity-selects efficiently. The allyl derivatives also showed an increase in the level of specific binding to targeted sequences compared with 2′-O-methyl probes of identical sequence. These properties indicate that the 2′-O-allyl oligoribonucleotides are particularly well suited for use as antisense probes.

Original languageEnglish
Pages (from-to)7752-7756
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number19
Publication statusPublished - 1 Dec 1990

Keywords

  • Affinity selection
  • Biotinylation
  • Modified RNA
  • Oligonucleotide synthesis
  • Ribonucleoprotein complexes

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