Combinatorial metabolic engineering using an orthogonal tri-functional CRISPR system

Jiazhang Lian, Mohammad HamediRad, Sumeng Hu and Huimin Zhao ()
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Jiazhang Lian: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Mohammad HamediRad: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Sumeng Hu: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign
Huimin Zhao: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign

Nature Communications, 2017, vol. 8, issue 1, 1-9

Abstract: Abstract Designing an optimal microbial cell factory often requires overexpression, knock-down, and knock-out of multiple gene targets. Unfortunately, such rewiring of cellular metabolism is often carried out sequentially and with low throughput. Here, we report a combinatorial metabolic engineering strategy based on an orthogonal tri-functional CRISPR system that combines transcriptional activation, transcriptional interference, and gene deletion (CRISPR-AID) in the yeast Saccharomyces cerevisiae. This strategy enables perturbation of the metabolic and regulatory networks in a modular, parallel, and high-throughput manner. We demonstrate the application of CRISPR-AID not only to increase the production of β-carotene by 3-fold in a single step, but also to achieve 2.5-fold improvement in the display of an endoglucanase on the yeast surface by optimizing multiple metabolic engineering targets in a combinatorial manner.

Date: 2017
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DOI: 10.1038/s41467-017-01695-x

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