Wicket: A Versatile Tool for the Integration and Optimization of Exogenous Pathways in Saccharomyces cerevisiae

ACS Synthetic Biology
Sha HouJun-Biao Dai

Abstract

Yeast can be used as a microbial cell factory to produce valuable chemicals. However, introducing an exogenous pathway into particular or different chromosomal locations for stable expression is still a daunting task. To address this issue, we designed a DNA cassette called a "wicket", which can be integrated into the yeast genome at designated loci to accept exogenous DNA upon excision by a nuclease. Using this system, we demonstrated that, in strains with "wickets", we could achieve near 100% efficiency for integration of the β-carotene pathway with no need for selective markers. Furthermore, it allowed independent and simultaneous integration of different genes in a pathway, resulting in a large variety of strains with variable copy numbers of each gene. This system could be a useful tool to modulate the integration of multiple copies of genes within a metabolic pathway and to optimize the yield of the target products.

References

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Dec 23, 2011·FEMS Yeast Research·Il-Kwon KimJens Nielsen
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Nov 3, 2015·Biotechnology and Bioengineering·Eugene FletcherJens Nielsen
Mar 19, 2016·Journal of Cellular Physiology·Jessica M WalterAndrew A Horwitz

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Citations

Sep 21, 2019·Biotechnology and Applied Biochemistry·Monika Yadav, Pratyoosh Shukla
Nov 17, 2020·Frontiers in Bioengineering and Biotechnology·Koray MalcıLeonardo Rios-Solis
Apr 29, 2021·Applied Microbiology and Biotechnology·Zi-Xu ZhangHe Huang
Oct 30, 2018·ACS Synthetic Biology·Leanne BourgeoisVincent J J Martin

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