Potent and selective inhibition of A-to-I RNA editing with 2′-O-methyl/locked nucleic acid-containing antisense oligoribonucleotides

Rena A. Mizrahi, Nicole T. Schirle, Peter A. Beal

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

ADARs (adenosine deaminases acting on RNA) are RNA editing enzymes that bind double helical RNAs and deaminate select adenosines (A). The product inosine (I) is read during translation as guanosine (G), so such changes can alter codon meaning. ADAR-catalyzed A to I changes occur in coding sequences for several proteins of importance to the nervous system. However, these sites constitute only a very small fraction of known A to I sites in the human transcriptome, and the significance of editing at the vast majority sites is unknown at this time. Site-selective inhibitors of RNA editing are needed to advance our understanding of the function of editing at specific sites. Here we show that 2′-O-methyl/locked nucleic acid (LNA) mixmer antisense oligonucleotides are potent and selective inhibitors of RNA editing on two different target RNAs. These reagents are capable of binding with high affinity to RNA editing substrates and remodeling the secondary structure by a strand-invasion mechanism. The potency observed here for 2′-O-methyl/LNA mixmers suggests this backbone structure is superior to the morpholino backbone structure for inhibition of RNA editing. Finally, we demonstrate antisense inhibition of editing of the mRNA for the DNA repair glycosylase NEIL1 in cultured human cells, providing a new approach to exploring the link between RNA editing and the cellular response to oxidative DNA damage.

Original languageEnglish (US)
Pages (from-to)832-839
Number of pages8
JournalACS Chemical Biology
Volume8
Issue number4
DOIs
StatePublished - Apr 19 2013

Fingerprint

Antisense Oligoribonucleotides
RNA Editing
RNA
Adenosine Deaminase
DNA Glycosylases
Morpholinos
Inosine
Guanosine
Antisense Oligonucleotides
Transcriptome
Codon
DNA Repair
Adenosine
Nervous System
DNA Damage
immune RNA
locked nucleic acid
Cultured Cells
DNA
Neurology

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Potent and selective inhibition of A-to-I RNA editing with 2′-O-methyl/locked nucleic acid-containing antisense oligoribonucleotides. / Mizrahi, Rena A.; Schirle, Nicole T.; Beal, Peter A.

In: ACS Chemical Biology, Vol. 8, No. 4, 19.04.2013, p. 832-839.

Research output: Contribution to journalArticle

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