5 years ago

Automated multiplex genome-scale engineering in yeast

Automated multiplex genome-scale engineering in yeast
Ran Chao, Wen Ren, Tong Si, Yuhao Min, Yuying Wu, Huimin Zhao
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.

Publisher URL: http://www.nature.com/articles/ncomms15187

DOI: 10.1038/ncomms15187

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