Automated multiplex genome-scale engineering in yeast

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. Genome-scale engineering is a powerful technique for understanding biology and designing microorganisms but has been limited to bacterial species. Here the authors present an automated platform for genome-scale engineering inSaccharomyces cerevisiaeusing CRISPR-Cas and RNAi.

• Publication year

  2017

• Venue

  Nature Communications

• Publication date

  2017-05-04

• Fields of study

  Biology, Medicine, Engineering

• Identifiers
  DOI 10.1038/ncomms15187PMID 28469255PMCID 5418614
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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