Probing Adenosine-to-Inosine Editing Reactions Using RNA-Containing Nucleoside Analogs

Olena Maydanovych, LaHoma M. Easterwood, Tao Cui, Eduardo A. Véliz, Subhash Pokharel, Peter A. Beal

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Advances in chemical synthesis and characterization of nucleic acids allows for atom-specific modification of complex RNAs, such as present in RNA editing substrates. By preparing substrates for ADARs by chemical synthesis, it is possible to subtly alter the structure of the edited nucleotide. Evaluating the effect these changes have on the rate of enzyme-catalyzed deamination reveals features of the editing reaction and guides the design of inhibitors. We describe the synthesis of select nucleoside analog phosphoramidites and their incorporation into RNAs that mimic known editing sites by solid phase synthesis, and analyze the interaction of these synthetic RNAs with ADARs using deamination kinetics and quantitative gel mobility shift assays.

Original languageEnglish (US)
Pages (from-to)369-386
Number of pages18
JournalMethods in Enzymology
Volume424
DOIs
StatePublished - 2007
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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    Maydanovych, O., Easterwood, L. M., Cui, T., Véliz, E. A., Pokharel, S., & Beal, P. A. (2007). Probing Adenosine-to-Inosine Editing Reactions Using RNA-Containing Nucleoside Analogs. Methods in Enzymology, 424, 369-386. https://doi.org/10.1016/S0076-6879(07)24017-0