Liu Ling, Chen Zhen, Tian Xiwei, Chu Ju
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
Biotechnol Lett. 2022 Jun;44(5-6):755-766. doi: 10.1007/s10529-022-03255-w. Epub 2022 May 8.
The target sorB gene, related to sorbicillinoid production, and the free expression element, AMA1, were used to verify the methodological approach in Acremonium chrysogenum.
CRISPR-Cas9 episomal expression system was used to introduce a point mutation into the sorB gene and the addition of sorB donor DNA achieved complete knockout of target genes. Four BSSS (yeast bud site selection system)-related genes, axl1, axl2, bud3, and bud4 were knocked out without impact on yield, dry weight, or pH. Relationships between morphology and stress tolerance in knockout strains were analyzed.
The gene-editing system used in the current study exceeded 80% efficiency and arthrospores development was found to differ from that in wild-type strain.
利用与山梨醇类物质产生相关的目标sorB基因和游离表达元件AMA1，验证产黄顶孢霉中的方法。
采用CRISPR-Cas9附加型表达系统在sorB基因中引入点突变，添加sorB供体DNA实现了目标基因的完全敲除。敲除了四个与酵母芽位点选择系统（BSSS）相关的基因axl1、axl2、bud3和bud4，对产量、干重或pH值没有影响。分析了敲除菌株中形态与胁迫耐受性之间的关系。
本研究中使用的基因编辑系统效率超过80%，发现节孢子的发育与野生型菌株不同。
