Use of mRNA- and protein-destabilizing elements to develop a highly responsive reporter system

D.C. Voon, L.S. Subrata, S. Baltic, M.P. Leu, J.M. Whiteway, A. Wong, S.A. Knight, Frank Christiansen, J.M. Daly

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Abstract

Reporter assays are widely used in applications that require measurement of changes in gene expression over time (e.g. drug screening). With standard reporter vectors, the measurable effect of a treatment or compound (altered reporter activity) is substantially diluted and delayed, compared with its true effect (altered transcriptional activity). This problem is caused by the relatively long half-lives of both the reporter protein and its mRNA. As a result, the activities of compounds, ligands or treatments that have a relatively minor effect, or a substantial but transient effect, often remain undetected. To circumvent this problem, we introduced modular protein- and mRNA-destabilizing elements into a range of commonly used reporters. Our data show that both elements are required for maximal responses to both increases and decreases in transcriptional activity. The double-destabilized reporter vectors showed markedly improved performance in drug screening, kinetic assays and dose-response titrations.
Original languageEnglish
Pages (from-to)online - approx 5-20pp
JournalNucleic Acids Research
Volume33
Issue number3
DOIs
Publication statusPublished - 2005

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Voon, D. C., Subrata, L. S., Baltic, S., Leu, M. P., Whiteway, J. M., Wong, A., ... Daly, J. M. (2005). Use of mRNA- and protein-destabilizing elements to develop a highly responsive reporter system. Nucleic Acids Research, 33(3), online - approx 5-20pp. https://doi.org/10.1093/nar/gni030