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

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Wicket: A Versatile Tool for the Integration and Optimization of Exogenous Pathways in Saccharomyces cerevisiae
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-02-23 00:00:00 , DOI: 10.1021/acssynbio.7b00391
Sha Hou ₁ , Qin Qin ₁ , Junbiao Dai _{1,

2}

Affiliation

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.

中文翻译：

小门：一种多功能工具，用于整合和优化酿酒酵母中的外源途径。

酵母可用作微生物细胞工厂，以生产有价值的化学物质。然而，将外源途径引入特定或不同的染色体位置以稳定表达仍然是艰巨的任务。为了解决这个问题，我们设计了一个称为“ wicket”的DNA盒，可以将其整合到酵母基因组的指定位点上，以便在被核酸酶切除后接受外源DNA。使用该系统，我们证明了，在具有“小门”的菌株中，无需选择标记，即可实现近100％的β-胡萝卜素途径整合效率。此外，它允许将不同基因独立并同时整合到途径中，从而导致每个基因的拷贝数可变的多种菌株。

更新日期：2018-02-23

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