Liu Xiaoqin, Deng Jingyu, Zhang Jinhong, Cui Zhiyong, Qi Qingsheng, Hou Jin
State Key Laboratory of Microbial Technology, Shandong University, Binhai Road 72, Qingdao, 266237, Shandong, People's Republic of China.
Biotechnol Biofuels Bioprod. 2024 Feb 15;17(1):24. doi: 10.1186/s13068-024-02472-x.
Genome-scale screening can be applied to efficiently mine for unknown genes with phenotypes of interest or special functions. It is also useful to identify new targets for engineering desirable properties of cell factories.
Here, we designed a new approach for genome-scale transcription activation using non-homologous end joining (NHEJ)-mediated integration in Yarrowia lipolytica. We utilized this approach to screen for genes that, upon activation, confer phenotypes including improved acetic acid tolerance and xylose metabolism. The candidates were validated using gene overexpression, and functional changes including improved growth performance under multiple stressors and activated pentose metabolism were identified.
This study provides a simple and effective approach to randomly activate endogenous genes and mine for key targets associated with phenotypes of interest. The specific gene targets identified here will be useful for cell factory construction and biorefining lignocellulose.
基因组规模筛选可用于高效挖掘具有感兴趣表型或特殊功能的未知基因。它对于识别用于改造细胞工厂理想特性的新靶点也很有用。
在此，我们设计了一种在解脂耶氏酵母中使用非同源末端连接（NHEJ）介导的整合进行基因组规模转录激活的新方法。我们利用这种方法筛选激活后赋予包括提高乙酸耐受性和木糖代谢等表型的基因。通过基因过表达对候选基因进行了验证，并鉴定了包括在多种应激条件下改善生长性能和激活戊糖代谢在内的功能变化。
本研究提供了一种简单有效的方法来随机激活内源基因并挖掘与感兴趣表型相关的关键靶点。此处鉴定出的特定基因靶点将有助于细胞工厂构建和木质纤维素生物精炼。
