State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, China.
Microb Biotechnol. 2022 Aug;15(8):2223-2234. doi: 10.1111/1751-7915.14060. Epub 2022 Apr 18.
With the rapid development of synthetic biology, the oleaginous yeast Yarrowia lipolytica has become an attractive microorganism for chemical production. To better optimize and reroute metabolic pathways, we have expanded the CRISPR-based gene expression toolkit of Y. lipolytica. By sorting the integration sites associated with high expression, new neutral integration sites associated with high expression and high integration efficiency were identified. Diverse genetic components, including promoters and terminators, were also characterized to expand the expression range. We found that in addition to promoters, the newly characterized terminators exhibited large variations in gene expression. These genetic components and integration sites were then used to regulate genes involved in the lycopene biosynthesis pathway, and different levels of lycopene production were achieved. The CRISPR-based gene expression toolkit developed in this study will facilitate the genetic engineering of Y. lipolytica.
随着合成生物学的快速发展，产油酵母解脂耶氏酵母已成为化学物质生产的一种有吸引力的微生物。为了更好地优化和改道代谢途径，我们扩展了 Yarrowia lipolytica 的基于 CRISPR 的基因表达工具包。通过对与高表达相关的整合位点进行排序，鉴定出了新的与高表达和高整合效率相关的中性整合位点。还对各种遗传元件，包括启动子和终止子进行了表征，以扩展表达范围。我们发现，除了启动子之外，新表征的终止子在基因表达方面也有很大的差异。然后，这些遗传元件和整合位点被用于调节番茄红素生物合成途径中的基因，实现了不同水平的番茄红素生产。本研究中开发的基于 CRISPR 的基因表达工具包将有助于 Yarrowia lipolytica 的基因工程改造。
