Comparative hybrid arrest by tandem antisense oligodeoxyribonucleotides or oligodeoxyribonucleoside methylpbosphonates in a cell-free system

Louis J. Maher, Bruce J. Dolnick

Research output: Contribution to journalArticle

103 Scopus citations

Abstract

Antisense oligonucleotides containing either anionic diester or neutral methylphosphonate internucleoside linkages were prepared by automated synthesis, and were compared for their ability to arrest translation of human dihydrofolate reductase (DHFR) mRNA in a nuclease treated rabbit reoculocyte lysate. In the case of oUgodeoxyribonucleotides, tandem targeting of three 14-mers resulted in synergistic and complete selective inhibition of DHFR synthesis at a total oligomer concentration of 25 μM. Hybrid arrest by three or six tandem oligodeoxyribonucleoside methylpnosphonates was dramatically less effective. This difference does not result from preferential recognition of hybrids involving oligodeoxyribonucleotides by endogenous RNaseH activity. A ribonuclease protection assay demonstrated that antisense oligodeoxyribonucleoside methylphosphonates bind selectively to target RNA sequences, but with 275 fold lower affinity than the corresponding oligodeoxyribonucleotides. This low binding affinity results in poor arrest of translation, and may be related to the stereochemistry of the methylphospbonate linkage.

Original languageEnglish (US)
Pages (from-to)3341-3358
Number of pages18
JournalNucleic acids research
Volume16
Issue number8
DOIs
StatePublished - Apr 25 1988

ASJC Scopus subject areas

  • Genetics

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