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Studying Nonsense-Mediated mRNA Decay in Mammalian Cells Using a Multicolored Bioluminescence-Based Reporter System

Andrew Nickless1, Zhongsheng You2

  • 1Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 14, 2017
PubMed

Insights

Researchers developed a novel bioluminescence reporter to quickly assess nonsense-mediated mRNA decay (NMD) activity in mammalian cells. This tool aids research into NMD

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Nonsense-mediated mRNA decay (NMD) is a surveillance pathway that degrades aberrant transcripts with premature termination codons (PTCs).
  • NMD also regulates physiological mRNA levels and its activity varies across cell types and conditions.
  • Modulating NMD holds therapeutic potential for genetic diseases and cancers.

Purpose of the Study:

  • To develop a simple, rapid, and effective reporter system for assaying NMD activity in live mammalian cells.
  • To facilitate basic and translational research on the NMD pathway.

Main Methods:

  • Development of a multicolored bioluminescence-based reporter system.
  • Introduction of the reporter into mammalian cells (transiently or stably) and animals.
  • Assessment of NMD activity using bioluminescence imaging, western blot, or RT-qPCR.

Main Results:

  • The reporter system accurately and efficiently measures NMD activity.
  • The system is versatile, applicable to cultured cells and whole animals.
  • Provides a straightforward method for NMD assessment.

Conclusions:

  • The described bioluminescence reporter system offers a valuable tool for studying NMD.
  • This method simplifies and accelerates NMD activity assessment in various biological contexts.
  • Facilitates research towards therapeutic applications targeting NMD.
Keywords:
Bioluminescence imagingCBGCBRNMD reporterNonsense-mediated mRNA decay (NMD)Western blottingqPCR

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