ArticleFungal genetics and biology : FG & B2020
Development of a marker-free mutagenesis system using CRISPR-Cas9 in the pathogenic mould Aspergillus fumigatus.
Article in Fungal genetics and biology : FG & B, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- Accelerated mutator phenotype in a clinicalEmerging microbes & infections · 2026Article
- Amino acid substitutions at the G118 hotspot in dihydroorotate dehydrogenase confer olorofim resistance inMicrobiology spectrum · 2026Article
- Article
- A High-Efficiency CRISPR-Cas9 Ribonucleoprotein Genome Editing System inJournal of fungi (Basel, Switzerland) · 2026Article
- Cas9 Ribonucleoproteins (RNPs) for Gene Deletion.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Marker-Free CRISPR/Cas9 RNP Transformation for Gene Disruption in Aspergillus fumigatus.Methods in molecular biology (Clifton, N.J.) · 2026Article
- The sulfur-related metabolic status ofVirulence · 2025Article
- Simultaneous multigene integration in Aspergillus fumigatus using CRISPR/Cas9 and endogenous counter-selectable markers.Journal of biological engineering · 2025Article
- High-Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus.Current protocols · 2025Article
- Recent developments inMicrobiology and molecular biology reviews : MMBR · 2025Review
- Article
- Co-transformation of Aspergillus fumigatus: a simple and efficient strategy for gene editing without linking selectable markers.Access microbiology · 2025Article
- Elevated mutation rates in multi-azole resistant Aspergillus fumigatus drive rapid evolution of antifungal resistance.Nature communications · 2024Article
- Shining a light on the impact of antifungals onAntimicrobial agents and chemotherapy · 2024Article
- Functional analysis of the Aspergillus fumigatus kinome identifies a druggable DYRK kinase that regulates septal plugging.Nature communications · 2024Article
- The Narrow Footprint of Ancient Balancing Selection Revealed by Heterokaryon Incompatibility Genes in Aspergillus fumigatus.Molecular biology and evolution · 2024Article
- Aspergillus fumigatus Hypoxia Adaptation Is Critical for the Establishment of Fungal Keratitis.Investigative ophthalmology & visual science · 2024Article
- Aspergillus fumigatus strains that evolve resistance to the agrochemical fungicide ipflufenoquin in vitro are also resistant to olorofim.Nature microbiology · 2024Article
- The CmSphere · 2023Article
- Efficient Generation of Multiple Seamless Point Mutations Conferring Triazole Resistance inJournal of fungi (Basel, Switzerland) · 2023Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Aspergillus fumigatus is a saprophytic fungal pathogen that is the cause of more than 300,000 life-threatening infections annually. Our understanding of pathogenesis and factors contributing to disease progression are limited. Development of rapid and versatile gene editing methodologies for A. fumigatus is essential. CRISPR-Cas9 mediated transformation has been widely used as a novel genome editing tool and has been used for a variety of editing techniques, such as protein tagging, gene deletions and site-directed mutagenesis in A. fumigatus. However, successful genome editing relies on time consuming, multi-step cloning procedures paired with the use of selection markers, which can result in a metabolic burden for the host and/or unintended transcriptional modifications at the site of integration. We have used an in vitro CRISPR-Cas9 assembly methodology to perform selection-free genome editing, including epitope tagging of proteins and site-directed mutagenesis. The repair template used during this transformation use 50 bp micro-homology arms and can be generated with a single PCR reaction or by purchasing synthesised single stranded oligonucleotides, decreasing the time required for complex construct synthesis.
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