Antisense-mediated affinity purification of dengue virus ribonucleoprotein complexes from infected cells

Stacia L. Phillips, Mariano Garcia-Blanco, Shelton Bradrick

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

The identification of RNA-binding proteins that physically associate with viral RNA molecules during infection can provide insight into the molecular mechanisms of RNA virus replication. Until recently, such RNA-protein interactions have been identified predominantly with the use of in vitro assays that may not accurately reflect associations that occur in the context of a living cell. Here we describe a method for the specific affinity purification of dengue virus RNA and associated proteins using in vivo cross-linking followed by antisense-mediated affinity purification. RNA-binding proteins that specifically co-purify with viral RNA using this method can be identified en masse by mass spectrometry. This strategy can potentially be adapted to the purification of any viral RNA species of interest.

Original languageEnglish (US)
JournalMethods
DOIs
StateAccepted/In press - Jul 6 2015

Keywords

  • Dengue virus
  • Ribonucleoprotein complex
  • RNA affinity chromatography

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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