2'-O-alkyl oligoribonucleotides as antisense probes

A. M. Iribarren, B. S. Sproat, P. Neuner, I. Sulston, U. Ryder, A. I. Lamond

Research output: Contribution to journalArticle

103 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 nucleases. 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)7747-7751
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number19
DOIs
Publication statusPublished - 1 Oct 1990

Keywords

  • affinity selection
  • biotinylation
  • modified RNA
  • ribonucleoprotein complexes oligonucleotide synthesis

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