Monitoring Enzymatic RNA G-Quadruplex Unwinding Activities by Nuclease Sensitivity and Reverse Transcription Stop Assays

Ewan K.S. McRae, Steven J. Dupas, Negar Atefi, Sean A. McKenna

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Multiple different methods have been employed to investigate the unwinding of RNA G-quadruplexes by various helicase proteins. Each has their own pitfalls, namely, looking at non-native or chemically modified RNA sequences, biasing the unwinding process with competing trap nucleotides, and a lack of context sequence to the 5′ and 3′ of the RNA G-quadruplex structure. Herein we present two straightforward methods that allow for quadruplex unwinding to be monitored on native RNA sequences without the use of fluorescent modifications, specialized equipment, or trap nucleotides to be employed.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages163-173
Number of pages11
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2209
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Helicase
  • Quadruplex
  • RNA
  • Unwinding

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint

Dive into the research topics of 'Monitoring Enzymatic RNA G-Quadruplex Unwinding Activities by Nuclease Sensitivity and Reverse Transcription Stop Assays'. Together they form a unique fingerprint.

Cite this