ArticleMicrobial biotechnology2024
Next-generation AMA1-based plasmids for enhanced heterologous expression in filamentous fungi.
Article in Microbial biotechnology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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4 citing papers in PubMed.
- Advances in molecular tools for elucidating nucleic acid biology in fungal pathogens.microLife · 2026Review
- Establishment of an antimetabolite-based transformation system for the wood-decaying basidiomyceteApplied and environmental microbiology · 2025Article
- Engineering of Global Transcriptional Regulators (GTRs) inJournal of fungi (Basel, Switzerland) · 2025Review
- Next-generation AMA1-based plasmids for enhanced heterologous expression in filamentous fungi.Microbial biotechnology · 2024Article
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7 authors.
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Abstract
Episomal AMA1-based plasmids are increasingly used for expressing biosynthetic pathways and CRISPR/Cas systems in filamentous fungi cell factories due to their high transformation efficiency and multicopy nature. However, the gene expression from AMA1 plasmids has been observed to be highly heterogeneous in growing mycelia. To overcome this limitation, here we developed next-generation AMA1-based plasmids that ensure homogeneous and strong expression. We achieved this by evaluating various degradation tags fused to the auxotrophic marker gene on the AMA1 plasmid, which introduces a more stringent selection pressure throughout multicellular fungal growth. With these improved plasmids, we observed in Aspergillus nidulans a 5-fold increase in the expression of a fluorescent reporter, a doubling in the efficiency of a CRISPRa system for genome mining, and a up to a 10-fold increase in the production of heterologous natural product metabolites. This strategy has the potential to be applied to diverse filamentous fungi.
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