Objective
The sorB gene related to sorbicillinoid production was used as the target, and the free expression element AMA1 was used to verify the availability of using this element in Acremonium chrysogenum.
Result
The point mutation of the sorB gene was successfully achieved in the Cripsr-Cas9 episomal expression system. In addition, the addition of sorB donor DNA in this system could achieve efficient, markless, and complete knockout of genes. Using this gene-editing platform, four BSSS-related genes Δaxl1, Δaxl2, Δbud3, and Δbud4 were knocked out completely, and it was found that the yield, dry weight, and pH of the knockout strains did not change significantly. Further, the stress tolerance of the knockout strains was determined, and the relationship between morphology and stress tolerance was preliminarily analyzed.
Conclusion
The gene-editing efficiency exceeded 80% and the developmental process of arthrospores differed from the starting strain.