ArticleScientific reports2019
Single crossover-mediated targeted nucleotide substitution and knock-in strategies with CRISPR/Cas9 system in the rice blast fungus.
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 9 papers.
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Who cites it
9 citing papers in PubMed, 35 citations in OpenAlex.
- Extracting Protoplasts from Filamentous Fungi Using Extralyse, An Enzyme Used in the Wine Industry.Current protocols · 2025Article
- The potential of genome editing to create novel alleles of resistance genes in rice.Frontiers in genome editing · 2024Review
- Article
- Establishment of a CRISPR/Cas9-Mediated Efficient Knockout System ofJournal of fungi (Basel, Switzerland) · 2023Article
- Genome editing using a versatile vector-based CRISPR/Cas9 system in Fusarium species.Scientific reports · 2022Article
- Calcium-based nanomaterials and their interrelation with chitosan: optimization for pCRISPR delivery.Journal of nanostructure in chemistry · 2022Article
- ZnAl nano layered double hydroxides for dual functional CRISPR/Cas9 delivery and enhanced green fluorescence protein biosensor.Scientific reports · 2020Article
- Plasmid-based and -free methods using CRISPR/Cas9 system for replacement of targeted genes in Colletotrichum sansevieriae.Scientific reports · 2019Article
- Progress and Challenges: Development and Implementation of CRISPR/Cas9 Technology in Filamentous Fungi.Computational and structural biotechnology journal · 2019Review
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome editing has become a promising approach for efficient and versatile genetic engineering in various organisms; however, simple and precise nucleotide modification methods in filamentous fungi have been restricted to double crossover type homologous recombination (HR). In this study, we developed a novel genome editing strategy via single crossover-mediated HR in the model filamentous fungus Pyricularia (Magnaporthe) oryzae. This method includes the CRISPR/Cas9 system and a donor vector harboring a single homology arm with point mutations at the CRISPR/Cas9 cleavage site. Using this strategy, we demonstrated highly efficient and freely programmable base substitutions within the desired genomic locus, and target gene disrupted mutants were also obtained via a shortened (100-1000 bp) single homology arm. We further demonstrated that this method allowed a one-step GFP gene knock-in at the C-terminus of the targeted gene. Since the genomic recombination does not require an intact protospacer-adjacent motif within the donor construct and any additional modifications of host components, this method can be used in various filamentous fungi for CRISPR/Cas9-based basic and applied biological analyses.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.