Establishment of a selectable marker recycling system for iterative gene editing in Fusarium fujikuroi

作     者：Tian-Qiong Shi Cai-Ling Yang Dong-Xun Li Yue-Tong Wang Zhi-Kui Nie 

作者机构：Jiangxi New Reyphon Biochemical Co.LtdSalt&Chemical IndustryXinganJiangxi331399People’s Republic of China School of Food Science and Pharmaceutical EngineeringNanjing Normal University2 Xuelin RoadQixia DistrictNanjing210023People’s Republic of China 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2024年第9卷第1期

页      面：159-164页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：supported by National key research and development program of China(2021YFC2104300) Natural Science Foundation of Jiangsu Province(BK20210573,20230381) Project of Leading Innovative Talents in Short-term Jiangxi Province(jxsq2023102172) Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project TSBICIP-PTJS-003-04 Postgraduate Research&Practice Innovation Program of Jiangsu Province KYCX23_1788 

主　　题：F.fujikuroi Gibberellic acid Plant growth hormone Selection marker recycling system CRISPR/Cas9 pyr4-blaster 

摘      要：Gibberellic acid(GA3)is a vital plant growth hormone widely used in ***,GA3 production relies on liquid fermentation by the filamentous fungus Fusarium ***,the lack of an effective selection marker recycling system hampers the application of metabolic engineering technology in ***,as multiple-gene editing and positive-strain screening still rely on a limited number of *** this study,we developed a strategy using pyr4-blaster and CRISPR/Cas9 tools for recycling orotidine-5 -phosphate decarboxylase(Pyr4)selection *** demonstrated the effectiveness of this method for iterative gene integration and large gene-cluster *** also successfully improved GA3 titers by overexpressing geranylgeranyl pyrophosphate synthase and truncated 3-hydroxy-3-methyl glutaryl coenzyme A reductase,which rewired the GA3 biosynthesis *** results highlight the efficiency of our established system in recycling selection markers during iterative gene editing ***,the selection marker recycling system lays the foundation for further research on metabolic engineering for GA3 industrial production.