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• W2900599367 abstract "Multiple gene modifications of specific fungal strains are required for achieving industrial-scale production of enzymes and secondary metabolites. In the present study, we developed an efficient multiple genetic engineering technique for the filamentous fungus Aspergillus oryzae . The approach is based on a clustered regulatory interspaced short palindromic repeats (CRISPR)/Cas9 system and recycling of an AMA1-based autonomous replicating plasmid. Because the plasmid harbors a drug resistance marker ( ptrA ), the approach does not require the construction of auxotrophic industrial strains prior to genome editing and allows for forced recycling of the gene-editing plasmid. The established plasmid-recycling technique involves an Aoace2 -conditional expression cassette, whose induction severely impairs fungal growth. We used the developed genetic engineering techniques for highly efficient marker-free multiple gene deletion/integration in A. oryzae . The genome-editing approaches established in the present study, which enable unlimited repeatable genetic engineering, will facilitate multiple gene modification of industrially important fungal strains." @default.
• W2900599367 created "2018-11-29" @default.
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• W2900599367 date "2019-02-01" @default.
• W2900599367 modified "2023-10-17" @default.
• W2900599367 title "Forced Recycling of an AMA1-Based Genome-Editing Plasmid Allows for Efficient Multiple Gene Deletion/Integration in the Industrial Filamentous Fungus <i>Aspergillus oryzae</i>" @default.
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• W2900599367 doi "https://doi.org/10.1128/aem.01896-18" @default.
• W2900599367 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6344613" @default.
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• W2900599367 hasPublicationYear "2019" @default.
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