期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 188, 期 4, 页码 1134-1144出版社
SPRINGER
DOI: 10.1007/s12010-019-02960-z
关键词
Acremonium chrysogenum; CRISPR; Cas9; Genome editing; Sorbicillinoid
资金
- NOW-MoST Joint Program [2013DFG32630]
- National Basic Research Program (973 program) [2012CB721006]
Acremonium chrysogenum is an important fungal strain used for cephalosporin C production. Many efforts have been made to develop versatile genome-editing tools to better understand the mechanism of A. chrysogenum. Here, we developed a feasible and efficient CRISPR/Cas9 system. Two genes responsible for the synthesis of yellow pigments (sorbicillinoids) were chosen as targets, and plasmids expressing both the Cas9 protein and single-guide RNAs were constructed. After introducing the plasmids into the protoplasts of A. chrysogenum, 83 to 93% albino mutants harboring the expected genomic alteration, on average, were obtained. We have generated two mutant strains that respectively disrupt sorA and sorB by flexible CRISPR/Cas9 system. We further confirmed that the sorbicillinoid biosynthetic gene cluster is regulated by an autoinduction mechanism. This work will lay a solid foundation for gene function research and regulation in the sorbicillinoid biosynthetic pathway.
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