Studying nonsense-mediated mRNA decay in mammalian cells using a multicolored bioluminescence-based reporter system

Andrew Nickless, Zhongsheng You

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

The nonsense-mediated mRNA decay (NMD) pathway degrades aberrant transcripts containing premature translation termination codons (PTCs) and also regulates the levels of many normal mRNAs containing NMD-inducing features. The activity of this pathway varies considerably in different cell types and can change in response to developmental and environmental cues. Modulating NMD activity represents a potential therapeutic avenue for certain genetic disorders and cancers. Simple reporter systems capable of faithfully assessing NMD activity in mammalian cells greatly facilitate both basic and translational research on NMD. Here we describe a simple and effective method for assaying NMD specifically and quickly in live mammalian cells using a multicolored bioluminescence-based reporter system. This reporter can be transiently or stably introduced into cultured cells as well as animals, and NMD activity can be accurately assessed by bioluminescence imaging, western blot, or RT-qPCR.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages213-224
Number of pages12
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1720
ISSN (Print)1064-3745

Keywords

  • Bioluminescence imaging
  • CBG
  • CBR
  • NMD reporter
  • Nonsense-mediated mRNA decay (NMD)
  • Western blotting
  • qPCR

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