Chapter title |
Studying Nonsense-Mediated mRNA Decay in Mammalian Cells Using a Multicolored Bioluminescence-Based Reporter System
|
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Chapter number | 16 |
Book title |
mRNA Decay
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Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7540-2_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7539-6, 978-1-4939-7540-2
|
Authors |
Andrew Nickless, Zhongsheng You |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 10 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 3 | 30% |
Researcher | 2 | 20% |
Unknown | 5 | 50% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 4 | 40% |
Design | 1 | 10% |
Unknown | 5 | 50% |