2′-O-alkyl oligoribonucleotides as antisense probes

TY - JOUR

T1 - 2′-O-alkyl oligoribonucleotides as antisense probes

AU - Iribarren, Adolfo M.

AU - Sproat, Brian S.

AU - Neuner, Philippe

AU - Sulston, Ingrid

AU - Ryder, Ursula

AU - Lamond, Angus I.

PY - 1990/12/1

Y1 - 1990/12/1

N2 - 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.

AB - 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.

KW - Affinity selection

KW - Biotinylation

KW - Modified RNA

KW - Oligonucleotide synthesis

KW - Ribonucleoprotein complexes

UR - https://www.scopus.com/pages/publications/11944259303

M3 - Article

AN - SCOPUS:11944259303

SN - 0027-8424

VL - 87

SP - 7752

EP - 7756

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 19

ER -