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Recent advances in the application of multiplex genome editing in Saccharomyces cerevisiae

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Abstract

Saccharomyces cerevisiae is a widely used microorganism and a greatly popular cell factory for the production of various chemicals. In order to improve the yield of target chemicals, it is often necessary to increase the copy numbers of key genes or engineer the related metabolic pathways, which traditionally required time-consuming repetitive rounds of gene editing. With the development of gene-editing technologies such as meganucleases, TALENs, and the CRISPR/Cas system, multiplex genome editing has entered a period of rapid development to speed up cell factory optimization. Multi-copy insertion and removing bottlenecks in biosynthetic pathways can be achieved through gene integration and knockout, for which multiplexing can be accomplished by targeting repetitive sequences and multiple sites, respectively. Importantly, the development of the CRISPR/Cas system has greatly increased the speed and efficiency of multiplex editing. In this review, the various multiplex genome editing technologies in S. cerevisiae were summarized, and the principles, advantages, and the disadvantages were analyzed and discussed. Finally, the practical applications and future prospects of multiplex genome editing were discussed.

Key points

• The development of multiplex genome editing in S. cerevisiae was summarized.

• The pros and cons of various multiplex genome editing technologies are discussed.

• Further prospects on the improvement of multiplex genome editing are proposed.

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Funding

This work was supported by the Nature Science Foundation of Jiangsu Province (No. BK20190706, No. BK20170988), the National Natural Science Foundation of China (No. 21908112, No. 22038007), National Key Research and Development Program of China (No.2019YFA0904900), and the Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project(TSBICIP-PTJS-003-04).

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Authors and Affiliations

  1. School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2 Xuelin Road, Qixia District, Nanjing, Jiangsu Province, China

    Zi-Xu Zhang, Ling-Ru Wang, Ying-Shuang Xu, Tian-Qiong Shi, Xiao-Man Sun & He Huang

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  1. Zi-Xu Zhang
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  2. Ling-Ru Wang
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  3. Ying-Shuang Xu
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  4. Wan-Ting Jiang
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  5. Tian-Qiong Shi
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  6. Xiao-Man Sun
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  7. He Huang
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Contributions

ZXZ, LRW, and WTJ wrote the original manuscript. YSX and TQS contributed the figures and table. XMS and HH revised the whole manuscript. All authors read and approved the manuscript.

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Correspondence to Tian-Qiong Shi or Xiao-Man Sun.

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The authors declare no competing interests.

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Zhang, ZX., Wang, LR., Xu, YS. et al. Recent advances in the application of multiplex genome editing in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 105, 3873–3882 (2021). https://doi.org/10.1007/s00253-021-11287-x

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