Abstract
The ability to perform complex bioassays in parallel enables experiments that are otherwise impossible because of throughput and cost constraints. For example, highly parallel chemical-genetic screens using pooled collections of thousands of defined Saccharomyces cerevisiae gene deletion strains are feasible because each strain is bar-coded with unique DNA sequences. It is, however, time-consuming and expensive to individually bar-code individual strains. To provide a simple and general method of barcoding yeast collections, we built a set of donor strains, called Barcoders, with unique bar codes that can be systematically transferred to any S. cerevisiae collection. We applied this technology by generating a collection of bar-coded 'decreased abundance by mRNA perturbation' (DAmP) loss-of-function strains comprising 87.1% of all essential yeast genes. These experiments validate both the Barcoders and the DAmP strain collection as useful tools for genome-wide chemical-genetic assays.
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Acknowledgements
We thank J. Horecka, N. Berbenetz and M. Urbanus for comments on the manuscript. This work is supported by grants from the US National Institute of Health and Canadian Institutes of Health Research to G.G. (MOP-81340) and to C.N. (MOP-84305), and from Genome Canada (to C.B. and B.J.A.).
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Z.Y. performed all bar-coding experiments and analysis and wrote this paper. M.C., B.J.A., J.P. and C.B. designed and created the original DAmP collection. L.E.H. did the data analysis. F.K. sequenced all bar codes. G.G. and C.N. designed the study, analyzed the data and wrote the paper.
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F.K. is employed by Prognosys Biosciences, Inc.
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Yan, Z., Costanzo, M., Heisler, L. et al. Yeast Barcoders: a chemogenomic application of a universal donor-strain collection carrying bar-code identifiers. Nat Methods 5, 719–725 (2008). https://doi.org/10.1038/nmeth.1231
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DOI: https://doi.org/10.1038/nmeth.1231
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