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Automated multiplex genome-scale engineering in yeast

Author

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  • Tong Si

    (Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
    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
    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
    University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
    University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA
    University of Illinois at Urbana-Champaign)

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.

Suggested Citation

  • Tong Si & Ran Chao & Yuhao Min & Yuying Wu & Wen Ren & Huimin Zhao, 2017. "Automated multiplex genome-scale engineering in yeast," Nature Communications, Nature, vol. 8(1), pages 1-12, August.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms15187
    DOI: 10.1038/ncomms15187
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    Cited by:

    1. Liu, Zihe & Moradi, Hamideh & Shi, Shuobo & Darvishi, Farshad, 2021. "Yeasts as microbial cell factories for sustainable production of biofuels," Renewable and Sustainable Energy Reviews, Elsevier, vol. 143(C).
    2. Dixit, Yatika & Yadav, Preeti & Sharma, Arun Kumar & Pandey, Poornima & Kuila, Arindam, 2023. "Multiplex genome editing to construct cellulase engineered Saccharomyces cerevisiae for ethanol production from cellulosic biomass," Renewable and Sustainable Energy Reviews, Elsevier, vol. 187(C).
    3. Siwei Li & Jingjing An & Yaqiu Li & Xiagu Zhu & Dongdong Zhao & Lixian Wang & Yonghui Sun & Yuanzhao Yang & Changhao Bi & Xueli Zhang & Meng Wang, 2022. "Automated high-throughput genome editing platform with an AI learning in situ prediction model," Nature Communications, Nature, vol. 13(1), pages 1-11, December.

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