An assessment of the utility of the yeast GreenScreen assay in pharmaceutical screening

Abstract

In this paper we describe an initial reproducibility study of 12 proprietary compounds followed by the assessment of 51 marketed pharmaceuticals and, lastly, a summary of the data so far from 2698 proprietary compounds from the Johnson & Johnson (J&J) compound library, in the yeast GreenScreen assay (GSA). In this assay, a reporter system in the yeast cells employs the DNA damage inducible promoter of the RAD54 gene, fused to the extremely stable green fluorescent protein (GFP). The assay proved to be very robust, the Excel templates provided by Gentronix with the assay interfaced well with in-house J&J systems with little adaptation, the assay was very rapid to perform and used very little compound. The results confirm previous work which suggests that the yeast GSA detects different classes of genotoxic compounds to the Ames assay and as a result can help screen out important genotoxic compounds at the pre-regulatory test phase that are missed by Ames-test-based screens alone. A combination of SAR evaluation of genotoxicity plus an Ames-test-based screen and the GSA provides a powerful pre-regulatory test battery to aid in the selection of successful drug candidates.

Cell-based in vitro models in environmental toxicology: a review

“…The UDS test can also be applied in vivo. A yeast-based fluorescence reporter assay for genotoxicity, termed GreenScreen, was introduced especially for application in high-throughput test systems [96]. The endpoint of the test represents chromosomeand DNA-metabolizing enzymes of yeast, as indicated by GFP-labeled expression reporter.…”

Genetic Toxicity:In VitroApproaches for Medicinal Chemists

“…This assay may be developed into a rather high-throughput test: one study reported screening 2698 compounds in a little more than a year [40]. An interlaboratory "ring trial" with 19 coded compounds concluded that the method provides an effective reproducible and reliable higher throughput screening alternative to the standard Ames test [41].…”

A pilot study for construction of a new cadmium-sensing yeast strain carrying a reporter plasmid with the <i>JLP1</i> promoter

“…The promoter sequences of RAD51, RAD54, HUG1, RNR2, and RNR3 have been used in reporter plasmids to detect agents that damage DNA, and assay strains have been shown to respond to various genotoxic agents such as gamma ray, methyl methanesulfonate, 4-nitroquinoline-1-oxide, methyl-Nnitro-N-nitrosoguanidine, cisplatin, and camptothecin (Walmsley et al, 1997;Billinton et al, 1998;Afanassiev et al, 2000;Boronat and Piña, 2006;Benton et al, 2007;Liu et al, 2008;Wei et al, 2013). The RAD54-GFP reporter assay strain has been validated as a genotoxicity test called the GreenScreen Assay (Cahill et al, 2004;Walsh et al, 2005;Van Gompel et al, 2005;Knight et al, 2007). The promoter sequences of the CUP1, SEO1, and UFO1 genes have been used to detect heavy metals.…”