ArticleScientific reports2019
High-throughput targeted gene deletion in the model mushroom Schizophyllum commune using pre-assembled Cas9 ribonucleoproteins.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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50 citing papers in PubMed, 83 citations in OpenAlex.
- Within-Population Genetic Structuring of the Cosmopolitan FungusEcology and evolution · 2026Article
- A High-Efficiency CRISPR-Cas9 Ribonucleoprotein Genome Editing System inJournal of fungi (Basel, Switzerland) · 2026Article
- The role of latitude and the Carpathian Mountain barrier in the spatial spread and population bottlenecks of Schizophyllum commune.BMC microbiology · 2026Article
- Effect of disruption of PsPCP gene on fruit body color ofMycoscience · 2026Article
- Gradients of Aliveness and Engineering: A Taxonomy of Fungal Engineered Living Materials.Advanced materials (Deerfield Beach, Fla.) · 2026Article
- Targeted insertion of heterogenous DNA using Cas9-gRNA ribonucleoprotein-mediated gene editing inBioengineered · 2025Article
- Strongest untreated mycelium materials produced by Schizophyllum commune dikaryons.World journal of microbiology & biotechnology · 2025Article
- CRISPR/Cas9-mediated genome editing in Ganoderma lucidum: recent advances and biotechnological opportunities.World journal of microbiology & biotechnology · 2025Review
- Development of a CRISPR/Cas9 RNP-mediated genetic engineering system in Paecilomyces variotii.Journal of microbiology (Seoul, Korea) · 2025Article
- The local-scale populations reveal cryptic processes occurring in the general population of Schizophyllum commune fungus.Scientific reports · 2025Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Review on mushroom mycelium-based products and their production process: from upstream to downstream.Bioresources and bioprocessing · 2025Review
- Simultaneous gene editing of both nuclei in a dikaryotic strain of Ganoderma lucidum using Cas9-gRNA ribonucleoprotein.Journal of microbiology (Seoul, Korea) · 2025Article
- Comparative transcriptome analyses and CRISPR/Cas9-mediated functional study ofFrontiers in microbiology · 2025Article
- Towards engineering agaricomycete fungi for terpenoid production.Journal of industrial microbiology & biotechnology · 2024Review
- Effect of a Mating Type Gene Editing inJournal of fungi (Basel, Switzerland) · 2024Article
- Unlocking Fungal Potential: The CRISPR-Cas System as a Strategy for Secondary Metabolite Discovery.Journal of fungi (Basel, Switzerland) · 2024Review
- Advancements in genetic studies of mushrooms: a comprehensive review.World journal of microbiology & biotechnology · 2024Review
- Current Advances in the Functional Genes of Edible and Medicinal Fungi: Research Techniques, Functional Analysis, and Prospects.Journal of fungi (Basel, Switzerland) · 2024Review
- Edible mycelium bioengineered for enhanced nutritional value and sensory appeal using a modular synthetic biology toolkit.Nature communications · 2024Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Efficient gene deletion methods are essential for the high-throughput study of gene function. Compared to most ascomycete model systems, gene deletion is more laborious in mushroom-forming basidiomycetes due to the relatively low incidence of homologous recombination (HR) and relatively high incidence of non-homologous end-joining (NHEJ). Here, we describe the use of pre-assembled Cas9-sgRNA ribonucleoproteins (RNPs) to efficiently delete the homeodomain transcription factor gene hom2 in the mushroom-forming basidiomycete Schizophyllum commune by replacing it with a selectable marker. All components (Cas9 protein, sgRNA, and repair template with selectable marker) were supplied to wild type protoplasts by PEG-mediated transformation, abolishing the need to optimize the expression of cas9 and sgRNAs. A Δku80 background further increased the efficiency of gene deletion. A repair template with homology arms of 250 bp was sufficient to efficiently induce homologous recombination. This is the first report of the use of pre-assembled Cas9 RNPs in a mushroom-forming basidiomycete and this approach may also improve the genetic accessibility of non-model species.
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