Nonsense-mediated mRNA decay (NMD) is a mechanism of mRNA surveillance ubiquitous among eukaryotes. Importantly, NMD not only removes aberrant transcripts with premature stop codons, but also regulates expression of many normal genes. A recently introduced dual-color fluorescent protein-based reporter enables analysis of NMD activity in live cells. In this chapter we describe the method to generate stable transgenic cell lines expressing the splicing-dependent NMD reporter using consecutive steps of lentivirus transduction and Tol2 transposition.
CITATION STYLE
Markina, N. M., Pereverzev, A. P., Staroverov, D. B., Lukyanov, K. A., & Gurskaya, N. G. (2018). Generation of cell lines stably expressing a fluorescent reporter of nonsense-mediated mRNA decay activity. In Methods in Molecular Biology (Vol. 1720, pp. 187–204). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7540-2_14
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