In vitro RNA cleavage assay for argonaute-family proteins

Keita Miyoshi, Hiroshi Uejima, Tomoko Nagami-Okada, Haruhiko Siomi, Mikiko C. Siomi

Research output: Chapter in Book/Report/Conference proceedingChapter

26 Citations (Scopus)


Recent studies have revealed that Argonaute proteins are crucial components of the RNA-induced silencing complexes (RISCs) that direct both small interfering RNA (siRNA)- and microRNA (miRNA)-mediated gene silencing. Full complementarity between the small RNA and its target messenger RNA (mRNA) results in RISC-mediated cleavage ("Slicing") of the target mRNA. A subset of Argonaute proteins directly contributes to the target cleavage ("Slicer") activity of the RISC. We describe (in vitro) Slicer assays using endogenous Argonaute protein immunopurified from animal cells and recombinant Argonaute protein produced in and purified from Escherichia coli.

Original languageEnglish
Title of host publicationRNAi
Subtitle of host publicationDesign and Application
PublisherHumana Press
Number of pages15
ISBN (Print)9781588298744
Publication statusPublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Argonaute
  • Drosophila
  • RISC
  • RNAi
  • Slicer

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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