Combinatorial metabolic engineering using an orthogonal tri-functional CRISPR system

Lian, Jiazhang; HamediRad, Mohammad; Hu, Sumeng; Zhao, Huimin

doi:10.1038/s41467-017-01695-x

Title: Combinatorial metabolic engineering using an orthogonal tri-functional CRISPR system

Journal Article · Wed Nov 22 00:00:00 EST 2017 · Nature Communications

DOI:https://doi.org/10.1038/s41467-017-01695-x· OSTI ID:1513269

^[1];

^[2]; Hu, Sumeng ^[2];

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Univ. of Illinois, Urbana, IL (United States); Zhejiang Univ., Hangzhou (China).
Univ. of Illinois, Urbana, IL (United States)

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 show that 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.

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Research Organization:: Univ. of Illinois at Urbana-Champaign, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018260

OSTI ID:: 1513269

Journal Information:: Nature Communications, Vol. 8, Issue 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 169 works

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