ArticleScientific reports2023
The Cas9-gRNA ribonucleoprotein complex-mediated editing of pyrG in Ganoderma lucidum and unexpected insertion of contaminated DNA fragments.
Article in Scientific reports, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed.
- Recent developments in CRISPR/Cas9 genome editing research for edible fungiculture.Functional & integrative genomics · 2026Review
- Screening of Monokaryotic Strains ofJournal of fungi (Basel, Switzerland) · 2025Article
- Targeted insertion of heterogenous DNA using Cas9-gRNA ribonucleoprotein-mediated gene editing inBioengineered · 2025Article
- Unraveling bioactive potential and production inChinese herbal medicines · 2025Review
- 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
- Gene Editing inJournal of fungi (Basel, Switzerland) · 2025Review
- Simultaneous gene editing of both nuclei in a dikaryotic strain of Ganoderma lucidum using Cas9-gRNA ribonucleoprotein.Journal of microbiology (Seoul, Korea) · 2025Article
- Towards engineering agaricomycete fungi for terpenoid production.Journal of industrial microbiology & biotechnology · 2024Review
- Trans-nuclei CRISPR/Cas9: safe approach for genome editing in the edible mushroom excluding foreign DNA sequences.Applied microbiology and biotechnology · 2024Article
- Effect of a Mating Type Gene Editing inJournal of fungi (Basel, Switzerland) · 2024Article
- Biosynthesis of iron-chelating terramides A-C and their role in Aspergillus terreus infection.Communications chemistry · 2024Article
- Advancements in genetic studies of mushrooms: a comprehensive review.World journal of microbiology & biotechnology · 2024Review
- Pleurotus ostreatus as a model mushroom in genetics, cell biology, and material sciences.Applied microbiology and biotechnology · 2024Review
- Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.Frontiers in microbiology · 2024Review
- An Efficient CRISPR/Cas9 Genome Editing System for aJournal of fungi (Basel, Switzerland) · 2023Article
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10 authors.
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Abstract
Gene editing is a promising alternative to traditional breeding for the generation of new mushroom strains. However, the current approach frequently uses Cas9-plasmid DNA to facilitate mushroom gene editing, which can leave residual foreign DNA in the chromosomal DNA raising concerns regarding genetically modified organisms. In this study, we successfully edited pyrG of Ganoderma lucidum using a preassembled Cas9-gRNA ribonucleoprotein complex, which primarily induced a double-strand break (DSB) at the fourth position prior to the protospacer adjacent motif. Of the 66 edited transformants, 42 had deletions ranging from a single base to large deletions of up to 796 bp, with 30 being a single base deletion. Interestingly, the remaining 24 contained inserted sequences with variable sizes at the DSB site that originated from the fragmented host mitochondrial DNA, E. coli chromosomal DNA, and the Cas9 expression vector DNA. The latter two were thought to be contaminated DNAs that were not removed during the purification process of the Cas9 protein. Despite this unexpected finding, the study demonstrated that editing G. lucidum genes using the Cas9-gRNA complex is achievable with comparable efficiency to the plasmid-mediated editing system.
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