ArticlePloS one2019
A new approach to Cas9-based genome editing in Aspergillus niger that is precise, efficient and selectable.
Article in PloS one, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- RoCi-a single-step multi-copy integration system based on rolling-circle replication.Nucleic acids research · 2026Article
- Unravelling fungal genome editing revolution: pathological and biotechnological application aspects.Archives of microbiology · 2025Review
- Unlocking Fungal Potential: The CRISPR-Cas System as a Strategy for Secondary Metabolite Discovery.Journal of fungi (Basel, Switzerland) · 2024Review
- Engineering of Aspergillus niger for efficient production of D-xylitol from L-arabinose.Microbial cell factories · 2024Article
- A novel luciferase-based reporter tool to monitor the dynamics of carbon catabolite repression in filamentous fungi.Microbial biotechnology · 2024Article
- Edible mycelium bioengineered for enhanced nutritional value and sensory appeal using a modular synthetic biology toolkit.Nature communications · 2024Article
- A Simple CRISPR/Cas9 System for Efficiently Targeting Genes of Aspergillus SectionMicrobiology spectrum · 2023Article
- Impact of overexpressing NADH kinase on glucoamylase production in Aspergillus niger.Journal of industrial microbiology & biotechnology · 2022Article
- CRISPR/Cas9-Based Genome Editing and Its Application inJournal of fungi (Basel, Switzerland) · 2022Review
- Article
- Advances in fungal chemical genomics for the discovery of new antifungal agents.Annals of the New York Academy of Sciences · 2021Review
- Development of a CRISPR/Cpf1 system for targeted gene disruption in Aspergillus aculeatus TBRC 277.BMC biotechnology · 2021Article
- CRISPR/Cas9-mediated genome editing in Penicillium oxalicum and Trichoderma reesei using 5S rRNA promoter-driven guide RNAs.Biotechnology letters · 2021Article
- Applications of CRISPR/Cas9 in the Synthesis of Secondary Metabolites in Filamentous Fungi.Frontiers in microbiology · 2021Review
- Protein expression and secretion by filamentous fungi.Journal of biosciences · 2021Review
- Genetic Manipulation and Transformation Methods forMycobiology · 2021Review
- Development of genetic tools for the thermophilic filamentous fungusBiotechnology for biofuels · 2020Article
- CRISPR/Cas9 genome editing technology in filamentous fungi: progress and perspective.Applied microbiology and biotechnology · 2019Review
- Upgrading of efficient and scalable CRISPR-Cas-mediated technology for genetic engineering in thermophilic fungusBiotechnology for biofuels · 2019Article
- Efficient marker free CRISPR/Cas9 genome editing for functional analysis of gene families in filamentous fungi.Fungal biology and biotechnology · 2019Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Aspergillus niger and other filamentous fungi are widely used in industry, but efficient genetic engineering of these hosts remains nascent. For example, while molecular genetic tools have been developed, including CRISPR/Cas9, facile genome engineering of A. niger remains challenging. To address these challenges, we have developed a simple Cas9-based gene targeting method that provides selectable, iterative, and ultimately marker-free generation of genomic deletions and insertions. This method leverages locus-specific "pop-out" recombination to suppress off-target integrations. We demonstrated the effectiveness of this method by targeting the phenotypic marker albA and validated it by targeting the glaA and mstC loci. After two selection steps, we observed 100% gene editing efficiency across all three loci. This method greatly reduces the effort required to engineer the A. niger genome and overcomes low Cas9 transformations efficiency by eliminating the need for extensive screening. This method represents a significant addition to the A. niger genome engineering toolbox and could be adapted for use in other organisms. It is expected that this method will impact several areas of industrial biotechnology, such as the development of new strains for the secretion of heterologous enzymes and the discovery and optimization of metabolic pathways.
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