Abstract:
:Nonsense-mediated mRNA decay (NMD) is an evolutionarily conserved mechanism of specific degradation of transcripts with a premature stop codon. NMD eliminates aberrant mRNAs arising from mutations, alternative splicing, and other events in cells. In addition, many normal transcripts undergo NMD. Recent studies demonstrated that NMD activity is specifically regulated and that NMD can play a role of global regulator of gene expression. Recently, we developed dual-color fluorescent protein-based reporters for quantification of NMD activity using fluorescence microscopy and flow cytometry (Pereverzev, Gurskaya, et al., 2015). Due to ratiometric fluorescence response, these reporters make it possible to assess NMD activity in live cells at the single-cell level and to reveal otherwise hidden heterogeneity of cells in respect of NMD activity. Here we provide a detailed description of applications of the NMD reporters in mammalian cell lines.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Gurskaya NG,Pereverzev AP,Staroverov DB,Markina NM,Lukyanov KAdoi
10.1016/bs.mie.2016.02.008subject
Has Abstractpub_date
2016-01-01 00:00:00pages
291-314eissn
0076-6879issn
1557-7988pii
S0076-6879(16)00092-6journal_volume
572pub_type
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