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

21 Citations (Scopus)

Abstract

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 publicationMethods in Molecular Biology
PublisherHumana Press
Pages29-43
Number of pages15
Volume442
ISBN (Print)9781588298744
DOIs
Publication statusPublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume442
ISSN (Print)10643745

Keywords

  • Argonaute
  • Drosophila
  • RISC
  • RNAi
  • Slicer

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Miyoshi, K., Uejima, H., Nagami-Okada, T., Siomi, H., & Siomi, M. C. (2008). In vitro RNA cleavage assay for argonaute-family proteins. In Methods in Molecular Biology (Vol. 442, pp. 29-43). (Methods in Molecular Biology; Vol. 442). Humana Press. https://doi.org/10.1007/978-1-59745-191-8_3