Detection of human Dicer and Argonaute 2 catalytic activity

Authors: Perron, MarjorieLandry, PatriciaPlante, IsabelleProvost, Patrick
Abstract: The microRNA (miRNA)-guided RNA silencing pathway is a central and well-defined cellular process involved in messenger RNA (mRNA) translational control. This complex regulatory process is achieved by a well orchestrated machinery composed of a relatively few protein components, among which the ribonuclease III (RNase III) Dicer and Argonaute 2 (Ago2) play a central role. These two proteins are essential and it is of particular interest to measure and detect their catalytic activity under various situations and/or conditions. In this chapter, we describe different protocols that aim to study and determine the catalytic activity of Dicer and Ago2 in cell extracts, immune complexes, and size-fractionated cell extracts. Another protocol aimed at assessing miRNA binding to Ago2 is also described. These experimental approaches are likely to be useful to researchers investigating the main steps of miRNA biogenesis and function in human health and diseases.
Document Type: Chapitre d'ouvrage
Issue Date: 5 April 2011
Open Access Date: Restricted access
Document version: VoR
Permalink: http://hdl.handle.net/20.500.11794/34270
This document was published in: Argonaute Proteins : Methods and Protocols ; Publié dans : Methods in Molecular Biology, Vol. 725, 121-141 (2011)
http://dx.doi.org/10.1007/978-1-61779-046-1_9
Springer
Alternative version: 10.1007/978-1-61779-046-1_9
21528451
Collection:Chapitres de livre

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