College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 South Puzhu Road, Nanjing, 211816, People's Republic of China.
Appl Microbiol Biotechnol. 2018 Nov;102(22):9541-9548. doi: 10.1007/s00253-018-9366-x. Epub 2018 Sep 21.
The oleaginous yeast Yarrowia lipolytica is widely used for the production of both bulk and fine chemicals, including organic acids, fatty acid-derived biofuels and chemicals, polyunsaturated fatty acids, single-cell proteins, terpenoids, and other valuable products. Consequently, it is becoming increasingly popular for metabolic engineering applications. Multiple gene manipulation tools including URA blast, Cre/LoxP, and transcription activator-like effector nucleases (TALENs) have been developed for metabolic engineering in Y. lipolytica. However, the low efficiency and time-consuming procedures involved in these methods hamper further research. The emergence of the CRISPR/Cas system offers a potential solution for these problems due to its high efficiency, ease of operation, and time savings, which can significantly accelerate the genomic engineering of Y. lipolytica. In this review, we summarize the research progress on the development of CRISPR/Cas systems for Y. lipolytica, including Cas9 proteins and sgRNA expression strategies, as well as gene knock-out/knock-in and repression/activation applications. Finally, the most promising and tantalizing future prospects in this area are highlighted.
油脂酵母解脂耶氏酵母被广泛用于生产大宗化学品和精细化学品，包括有机酸、脂肪酸衍生的生物燃料和化学品、多不饱和脂肪酸、单细胞蛋白、萜类化合物和其他有价值的产品。因此，它在代谢工程应用中越来越受欢迎。已经开发了多种基因操作工具，包括 URA 爆破、Cre/LoxP 和转录激活因子样效应物核酸酶（TALENs），用于解脂耶氏酵母的代谢工程。然而，这些方法涉及的低效率和耗时的程序阻碍了进一步的研究。CRISPR/Cas 系统的出现为这些问题提供了一个潜在的解决方案，因为它具有高效率、易于操作和节省时间的特点，这可以显著加速解脂耶氏酵母的基因组工程。在这篇综述中，我们总结了 CRISPR/Cas 系统在解脂耶氏酵母中的开发研究进展，包括 Cas9 蛋白和 sgRNA 表达策略，以及基因敲除/敲入和抑制/激活应用。最后，强调了该领域最有前途和诱人的未来前景。
