Dai Ziyu
Chemical and Biological Processes Development Group, Pacific Northwest National Laboratory, WA, Richland, USA.
Joint Bioenergy Institute, Emeryville, CA, United States.
FEBS J. 2023 Nov;290(21):5094-5097. doi: 10.1111/febs.16956. Epub 2023 Oct 4.
Since the first CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system was developed for creating double-stranded DNA breaks, it has been adapted and improved for different biotechnological applications. In this issue of The FEBS Journal, Arentshorst et al. developed a novel approach to enhance transgene expression of a specific protein, patulin synthase (PatE) from Penicillium expansum, in the important industrial filamentous fungus Aspergillus niger. Their technique involved the disruption of selected genes with counter-effects on targeted protein production and simultaneous integration of glucoamylase landing sites into the disrupted gene locus such as protease regulator (prtT) in an ATP-dependent DNA helicase II subunit 1 (kusA or ku70)-deletion strain. Multiple copies of the PatE transgene expression cassette were introduced by CRISPR-Cas9-mediated insertion. The purified PatE was further used for structural and functional studies, and the technique laid the foundation for elevating the overall production of various proteins or chemicals in those industrially important fungi.
自从首个用于产生双链DNA断裂的CRISPR（规律成簇的间隔短回文重复序列）-Cas（CRISPR相关蛋白）系统被开发以来，它已被调整和改进以用于不同的生物技术应用。在本期《FEBS杂志》中，阿伦肖斯特等人开发了一种新方法，以增强来自扩展青霉的特定蛋白质棒曲霉素合酶（PatE）在重要工业丝状真菌黑曲霉中的转基因表达。他们的技术包括破坏对目标蛋白质产生有反作用的选定基因，并将葡糖淀粉酶着陆位点同时整合到破坏的基因位点，如在ATP依赖性DNA解旋酶II亚基1（kusA或ku70）缺失菌株中的蛋白酶调节因子（prtT）。通过CRISPR-Cas9介导的插入引入了多个拷贝的PatE转基因表达盒。纯化后的PatE进一步用于结构和功能研究，该技术为提高那些工业重要真菌中各种蛋白质或化学品的总体产量奠定了基础。
