ArticleFungal genetics and biology : FG & B2018
Efficient genome editing in Fusarium oxysporum based on CRISPR/Cas9 ribonucleoprotein complexes.
Article in Fungal genetics and biology : FG & B, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.
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Who cites it
64 citing papers in PubMed, 152 citations in OpenAlex.
- A streamlined protocol for small-scale protoplast generation and CRISPR/Cpf1-mediated genome editing iniScience · 2026Article
- Establishment of an Efficient CRISPR-Cas9-Mediated Gene Disruption System in the Lichen-Forming FungusJournal of fungi (Basel, Switzerland) · 2026Article
- Metabolic Engineering for Gibberellic Acid Production inMolecules (Basel, Switzerland) · 2026Review
- The Effector SIX8 Contributes to Virulence of Fusarium oxysporum f. sp. lactucae Race 4 on Lettuce.Molecular plant pathology · 2026Article
- Soil Reservoirs of Antifungal-Resistant Fungi: Implications for Plant Disease Management with a Focus onMicroorganisms · 2026Review
- An ATMT-CRISPR/Cas9 system for genome editing in Monascus purpureus.World journal of microbiology & biotechnology · 2026Article
- The Multifaceted Menace ofBiology · 2026Review
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- Advances in molecular tools for elucidating nucleic acid biology in fungal pathogens.microLife · 2026Review
- Accessory Chromosome Contributes to Virulence of Banana Infecting Fusarium oxysporum Tropical Race 4.Molecular plant pathology · 2025Article
- Genetic and molecular approaches for Fusarium wilt resistance in garden pea: advances and future outlook.Plant molecular biology · 2025Review
- Regulation and induction of fungal secondary metabolites: a comprehensive review.Archives of microbiology · 2025Review
- Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi.Journal of fungi (Basel, Switzerland) · 2025Review
- Novel application of ribonucleoprotein-mediated CRISPR-Cas9 gene editing in plant pathogenic oomycete species.Microbiology spectrum · 2025Article
- Groundbreaking Technologies and the Biocontrol of Fungal Vascular Plant Pathogens.Journal of fungi (Basel, Switzerland) · 2025Review
- Optimization of a CRISPR-Cas9Frontiers in plant science · 2025Article
- CRISPR/Cas9-Engineering for Increased Amylolytic Potential of Microbes for Sustainable Wastewater Treatment: A Review.Current microbiology · 2024Review
- CRISPR-Cas9-mediated enhancement of Beauveria bassiana virulence with overproduction of oosporein.Fungal biology and biotechnology · 2024Article
- Unlocking Fungal Potential: The CRISPR-Cas System as a Strategy for Secondary Metabolite Discovery.Journal of fungi (Basel, Switzerland) · 2024Review
- A user-friendly CRISPR/Cas9 system for mutagenesis of Neurospora crassa.Scientific reports · 2024Article
4 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The Fusarium oxysporum species complex (FOSC) is an economically important group of pathogenic filamentous fungi that are able to infect both animals and plants. Reverse genetic techniques, including gene disruption/deletion methods, to study these fungi are available although limitations exist resulting in decreased efficiency. Herein we describe a gene editing system developed using a F. oxysporum-optimized Cas9 ribonucleoprotein (RNP) and protoplast transformation method. The Cas9 protein and sgRNA were assembled to form a stable RNP in vitro and this complex was transferred into fungal protoplasts for gene editing with PEG-mediated transformation. In order to determine if the Cas9 RNP system is functional in the FOSC protoplasts and assess the efficacy of the system, two genes, URA5 and URA3, were selected for targeted disruption generating uracil auxotroph mutants that are resistant to 5-fluoroorotic acid, 5-FOA. In addition, a gene in a secondary metabolite biosynthetic cluster, the ortholog of BIK1, was mutated using this system and the maximum efficiency of this gene disruption was about 50%. Further analysis of the bik1 mutant confirmed that this polyketide synthase was involved in the synthesis of the red pigment, bikaverin. The mutants generated in this study displayed the strong expected phenotypes, demonstrating this F. oxysporum-optimized CRISPR/Cas9 system is stable and can efficiently disrupt the genes of interest.
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