Automated multiplex genome-scale engineering in yeast

Tong Si, Ran Chao, Yuhao Min, Yuying Wu, Wen Ren and Huimin Zhao ()
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Tong Si: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
Ran Chao: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
Yuhao Min: University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
Yuying Wu: University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
Wen Ren: University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
Huimin Zhao: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA

Nature Communications, 2017, vol. 8, issue 1, 1-12

Abstract: Abstract Genome-scale engineering is indispensable in understanding and engineering microorganisms, but the current tools are mainly limited to bacterial systems. Here we report an automated platform for multiplex genome-scale engineering in Saccharomyces cerevisiae, an important eukaryotic model and widely used microbial cell factory. Standardized genetic parts encoding overexpression and knockdown mutations of >90% yeast genes are created in a single step from a full-length cDNA library. With the aid of CRISPR-Cas, these genetic parts are iteratively integrated into the repetitive genomic sequences in a modular manner using robotic automation. This system allows functional mapping and multiplex optimization on a genome scale for diverse phenotypes including cellulase expression, isobutanol production, glycerol utilization and acetic acid tolerance, and may greatly accelerate future genome-scale engineering endeavours in yeast.

Date: 2017
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DOI: 10.1038/ncomms15187

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