An assessment of the utility of the yeast GreenScreen assay in

0,11
MB pharmaceutical screening

6
stron

6619
ID University of Manchester

2005
rok

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 evaluati on 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.