Multi-step multi-mutant production method using genome editing in filamentous fungi

摘要

To provide a new technique for efficiently preparing Aspergillus oryzae strains having mutations in multiple genes using genome editing techniques.SOLUTION: The invention provides a genome editing plasmid vector comprising a gene encoding Cas9 nuclease, guiding RNA(s) for respective genome target site(s), a positive selection marker gene and a negative selection marker gene. The invention also provides a multi-step method for preparing multiplex mutants using the plasmid vector in genome editing in filamentous fungi.SELECTED DRAWING: Figure 4