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

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    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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