Assessment of the genotoxicity of S9-generated metabolites using the GreenScreen HC GADD45a-GFP assay

Christopher Jagger, Matthew Tate, Paul A. Cahill, Chris Hughes, Andrew W. Knight, Nicholas Billinton, Richard M. Walmsley

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Genotoxicity can be assessed by monitoring expression of a GADD45a-GFP reporter in the human lymphoblastoid cell line TK6. A flow cytometric method has been developed to effectively distinguish GFP fluorescence from coloured and fluorescent test samples as well from the S9 liver extracts used to generate metabolites from pro-genotoxins. The method includes the use of propidium iodide exclusion for the determination of cellular viability. This paper describes the method development, the derivation of decision thresholds for the identification of genotoxins using the method, and presents data from a 56-compound validation study of the method. The results illustrate that the method permitted the detection of the majority of pro-genotoxins tested and, importantly, the high specificity of the GADD45a-GFP assay was maintained. © The Author 2008. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved.
    Original languageEnglish
    Pages (from-to)35-50
    Number of pages15
    JournalMutagenesis
    Volume24
    Issue number1
    DOIs
    Publication statusPublished - Jan 2009

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