Heterokaryosis, the main obstacle in the generation of PPO1-edited Agaricus bisporus by CRISPR/Cas9 system

Article Forestry

Marker-free genome editing in the edible mushroom, Pleurotus ostreatus, using transient expression of genes required for CRISPR/Cas9 and for selection

Daishiro Koshi et al.

Summary: In this study, marker-free genome editing was achieved in Pleurotus ostreatus by combining CRISPR/Cas9 with a transient transformation system. This opens up possibilities for repeated genome editing in these fungi.

JOURNAL OF WOOD SCIENCE (2022)

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Article Multidisciplinary Sciences

Metabolomics and transcriptomics unravel the mechanism of browning resistance in Agaricus bisporus

Zhi-Xin Cai et al.

Summary: This study reveals the mechanism of browning in Agaricus bisporus and explores the differences between browning-resistant and easy-browning varieties. The results show that browning-resistant varieties have higher levels of organic acids, while easy-browning varieties have higher levels of trehalose, which may be important factors influencing browning. Furthermore, differential gene expression and metabolite levels also play a role in browning.

PLOS ONE (2022)

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Article Environmental Sciences

The Impact of Ambient Environmental and Occupational Pollution on Respiratory Diseases

Chinatsu Nishida et al.

Summary: Both ambient pollutants and occupational pollutants can cause and worsen lung and respiratory diseases. Particulate matter (PM2.5) is an important ambient pollutant, which can trigger asthma attacks and exacerbate chronic obstructive pulmonary disease in the short term, as well as increase the risk of lung cancer and mortality in the long term. Asbestos and crystalline silica are well-known traditional occupational pollutants leading to pneumoconiosis. Work-related asthma (WRA) has become the most common occupational lung disease in recent years, caused by various agents including natural and synthetic chemicals and irritant gases. Primary preventive interventions that raise awareness of pollutants and reduce the development and exacerbation of diseases caused by air pollutants are crucial in addressing both ambient and occupational pollution.

INTERNATIONAL JOURNAL OF ENVIRONMENTAL RESEARCH AND PUBLIC HEALTH (2022)

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Article Biotechnology & Applied Microbiology

CRISPR/Cas9 using a transient transformation system in Ceriporiopsis subvermispora

Takehito Nakazawa et al.

Summary: This study reports an efficient method for gene mutagenesis in Ceriporiopsis subvermispora using the CRISPR/Cas9 system. Results showed that the target gene(s) could be disrupted or modified successfully by introducing plasmids expressing Cas9 and single-guide RNA. Marker-free genome editing was observed in the mutants. This method provides a valuable tool for studying lignocellulose-degrading systems in C. subvermispora.

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY (2022)

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Article Biotechnology & Applied Microbiology

Construction of CRISPR-Cas9 genome editing platform for white-rot fungus Cerrena unicolor BBP6 and its effects on extracellular ligninolytic enzyme biosynthesis

Hao Zhang et al.

Summary: This study successfully developed a CRISPR-Cas9 genome editing platform for Cerrena unicolor BBP6. By disrupting specific genes, the laccase and MnP activities were reduced in the strain. This research is important for further understanding of ligninolytic enzyme biosynthesis in this fungus.

BIOCHEMICAL ENGINEERING JOURNAL (2022)

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Article Microbiology

Experimental Outcrossing in Agaricus bisporus Revealed a Major and Unexpected Involvement of Airborne Mycelium Fragments

Banafsheh Jalalzadeh et al.

Summary: Study reveals that button mushrooms can reproduce through both heterokaryotic and airborne mycelium fragment outcrossing, potentially explaining the high level of heterozygosity and cultivar introgression in wild populations.

JOURNAL OF FUNGI (2022)

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Article Biotechnology & Applied Microbiology

Efficient CRISPR/Cas9 system based on autonomously replicating plasmid with an AMA1 sequence and precisely targeted gene deletion in the edible fungus, Cordyceps militaris

Guoliang Meng et al.

Summary: In this study, an efficient CRISPR/Cas9 genome-editing system was successfully constructed in Cordyceps militaris, allowing for gene editing and precise gene deletion with high efficiencies. The findings demonstrate the potential of plasmid loss in avoiding adverse effects of the CRISPR/Cas9 system and facilitating the breeding of new cultivars. Additionally, RNA Pol III promoters, endogenous tRNA, and chimeric tRNA can be used to improve editing efficiency.

MICROBIAL BIOTECHNOLOGY (2022)

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Article Microbiology

Establishment of CRISPR/Cas9 Genome-Editing System Based on Dual sgRNAs in Flammulina filiformis

Xiaotian Liu et al.

Summary: This study successfully established a CRISPR/Cas9 genome editing system based on dual sgRNAs in Flammulina filiformis, allowing efficient genetic modifications and expanding the applications of CRISPR/Cas9 in Basidiomycetes.

JOURNAL OF FUNGI (2022)

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Article Biotechnology & Applied Microbiology

Multiplex gene precise editing and large DNA fragment deletion by the CRISPR-Cas9-TRAMA system in edible mushroom Cordyceps militaris

Bai-Xiong Chen et al.

Summary: In this study, a novel genomic editing system was developed to achieve precise gene editing and large synthetic cluster deletion in Cordyceps militaris. The system has important application value in exploring the metabolic resources of medicinal mushrooms and constructing cellular pathways.

MICROBIAL BIOTECHNOLOGY (2022)

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Letter Microbiology

Gene targeting of dikaryotic Pleurotus ostreatus nuclei using the CRISPR/Cas9 system

Fuga Yamasaki et al.

Summary: In this study, we reported a method for simultaneously targeting both nuclei of dikaryotic P. ostreatus PC9x#64 using plasmid-based CRISPR/Cas9, aiming to develop an efficient and rapid molecular breeding method. By targeting the meiosis-related genes mer3 and msh4, mutations were successfully introduced and expressed in both nuclei.

FEMS MICROBIOLOGY LETTERS (2022)

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Article Mycology

Molecular breeding of sporeless strains of Pleurotus ostreatus using a non-homologous DNA end-joining defective strain

Fuga Yamasaki et al.

Summary: In this study, gene targeting was used to disrupt msh4 and mer3 genes in Pleurotus ostreatus, resulting in changes in basidiospore production. Disruption of the ku80 gene did not significantly affect basidiospore production. This study represents the first report of molecular breeding of sporeless strains in cultivated mushrooms using an efficient method for targeted gene disruption.

MYCOLOGICAL PROGRESS (2021)

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Article Biotechnology & Applied Microbiology

Efficient genome editing with CRISPR/Cas9 in Pleurotus ostreatus

Tatpong Boontawon et al.

Summary: Efficient gene mutagenesis in Pleurotus ostreatus was achieved using plasmid-based CRISPR/Cas9, generating drug-resistant strains and introducing mutations via homology-directed repair. This approach could contribute to molecular breeding of non-GM cultivated strains in the future.

AMB EXPRESS (2021)

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Article Agronomy

Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes

Suyun Moon et al.

Summary: This study presents the first method for genetic engineering in shiitake mushrooms using CRISPR/Cas9, successfully disrupting target gene loci and affecting transcription levels of downstream genes.

MYCOBIOLOGY (2021)

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Article Genetics & Heredity

Generation of Iron-Independent Siderophore-Producing Agaricus bisporus through the Constitutive Expression of hapX

Min-Seek Kim et al.

Summary: Recombinant strain of Agaricus bisporus have been constructed to produce siderophore independently of iron by escaping hapX gene repression, showcasing a potential new application for mushrooms through molecular biological manipulation.

GENES (2021)

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Article Biotechnology & Applied Microbiology

Targeted Gene Insertion and Replacement in the Basidiomycete Ganoderma lucidum by Inactivation of Nonhomologous End Joining Using CRISPR/Cas9

Jun-Liang Tu et al.

Summary: This study successfully inactivated the nonhomologous end-joining (NHEJ) pathway in Ganoderma lucidum using the CRISPR/Cas9 system, by disrupting the Ku70 gene, to establish an efficient tool for targeted gene insertion and replacement. Strains with disrupted ku70 gene exhibited high gene targeting efficiency, advancing our understanding of the functional genomics of G. lucidum.

APPLIED AND ENVIRONMENTAL MICROBIOLOGY (2021)

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Article Microbiology

Gene targeting using pre-assembled Cas9 ribonucleoprotein and split-marker recombination in Pleurotus ostreatus

Tatpong Boontawon et al.

Summary: This study successfully utilized Cas9 RNP for gene mutagenesis in Pleurotus ostreatus, generating strains resistant to 5-fluoroorotic acid, and demonstrated its applicability for molecular breeding. Additionally, gene disruption via the Cas9 RNP system was also achieved in wild-type P. ostreatus, overcoming the challenges in traditional gene targeting studies.

FEMS MICROBIOLOGY LETTERS (2021)

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Article Biotechnology & Applied Microbiology