Methods to Analyze MicroRNA-Mediated Control of mRNA Translation

Jennifer L. Clancy, Marco Nousch, David T. Humphreys, Belinda J. Westman, Traude H. Beilharz, Thomas Preiss

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

40 Citations (Scopus)

Abstract

MicroRNAs (miRs) are an important class of gene regulators that affect a wide range of biological processes. Despite the early recognition of miRs as translational regulators and intense interest in studying this phenomenon, it has so far not been possible to derive a consensus model for the underlying molecular mechanism(s). The potential of miRs to act in a combinatorial manner and to also promote mRNA decay creates conceptual and technical challenges in their study. Here, we discuss critical parameters in design and analysis of experiments used to study miR function including creation of synthetic miR and mRNA partners for assay of translational inhibition using luciferase reporters; measurement of mRNA stability after miR action; defining poly(A) tail length in miR target mRNA; determining the distribution of miRs and their target mRNAs in polysome profiles; and visualization of P-body components. We describe protocols for each of these procedures.

Original languageEnglish
Title of host publicationTranslation Initiation
Subtitle of host publicationCell Biology, High Throughput Methods, and Chemical Based Approaches
Pages83-111
Number of pages29
Volume431
DOIs
Publication statusPublished - 2007
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume431
ISSN (Print)0076-6879

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