Title |
Two low complexity ultra-high throughput methods to identify diverse chemically bioactive molecules using Saccharomyces cerevisiae
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Published in |
Microbiological Research, February 2017
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DOI | 10.1016/j.micres.2017.02.004 |
Pubmed ID | |
Authors |
Katarina Petrovic, Martin Pfeifer, Christian N. Parker, Sven Schuierer, John Tallarico, Dominic Hoepfner, N. Rao Movva, Gunther Scheel, Stephen B. Helliwell |
Abstract |
The budding yeast S. cerevisiae is widely used as a eukaryotic model organism to elucidate the mechanism of action of low molecular weight compounds. This report describes the development of two high throughput screening methods based on cell viability either by monitoring the reduction of alamarBlue(®) (resazurin) or by direct optical measurement of cell growth. Both methods can be miniaturized to allow screening of large numbers of samples, and can be performed using S. cerevisiae in 384 and 1536-well format. The alamarBlue(®) approach achieves Z' values of >0.7 with signal to basal ratios of >6.5, and around 1.1 million low molecular weight compounds were screened, identifying approximately 25,000 primary hits. Dose response curves generated for a subset (1930) using both alamarBlue(®) and optical density methods showed significant overlap. In genome-wide haploinsufficiency profiling (HIP), 572 of these hits demonstrated a diverse mechanism of action, affecting >25% of all yeast strains. |
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France | 1 | 14% |
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Unknown | 4 | 57% |
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Student > Bachelor | 4 | 18% |
Student > Ph. D. Student | 4 | 18% |
Student > Master | 3 | 14% |
Other | 1 | 5% |
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Unknown | 2 | 9% |
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Neuroscience | 1 | 5% |
Other | 0 | 0% |
Unknown | 3 | 14% |