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

• DOI: 10.1093/mutage/gen050
• Publication Date: 2008-09-12T01:22:54Z
• Authors: C. Jagger; M. Tate; P. A. Cahill; C. Hughes; A. W. Knight; N. Billinton; R. M. Walmsley
• Keywords: Propidium iodide

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 S9 liver extracts used generate metabolites pro-genotoxins. The includes use propidium iodide exclusion for determination cellular viability. This paper describes development, derivation decision thresholds identification genotoxins using method, presents data 56-compound validation study method. results illustrate that permitted detection majority pro-genotoxins tested and, importantly, high specificity assay was maintained.