Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system
Abstract Mushroom-forming basidiomycetes produce a wide range of metabolites and have great value not only as food but also as an important global natural resource. Here, we demonstrate CRISPR/Cas9-based genome editing in the model species Coprinopsis cinerea. Using a high-throughput reporter assay...
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Nature Portfolio
2017
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oai:doaj.org-article:42f047196f1f4130a92caee3e78874202021-12-02T16:06:17ZGenome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system10.1038/s41598-017-00883-52045-2322https://doaj.org/article/42f047196f1f4130a92caee3e78874202017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00883-5https://doaj.org/toc/2045-2322Abstract Mushroom-forming basidiomycetes produce a wide range of metabolites and have great value not only as food but also as an important global natural resource. Here, we demonstrate CRISPR/Cas9-based genome editing in the model species Coprinopsis cinerea. Using a high-throughput reporter assay with cryopreserved protoplasts, we identified a novel promoter, CcDED1 pro , with seven times stronger activity in this assay than the conventional promoter GPD2. To develop highly efficient genome editing using CRISPR/Cas9 in C. cinerea, we used the CcDED1 pro to express Cas9 and a U6-snRNA promoter from C. cinerea to express gRNA. Finally, CRISPR/Cas9-mediated GFP mutagenesis was performed in a stable GFP expression line. Individual genome-edited lines were isolated, and loss of GFP function was detected in hyphae and fruiting body primordia. This novel method of high-throughput CRISPR/Cas9-based genome editing using cryopreserved protoplasts should be a powerful tool in the study of edible mushrooms.Shigeo S. SuganoHiroko SuzukiEisuke ShimokitaHirofumi ChibaSumihare NojiYuriko OsakabeKeishi OsakabeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017) |
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Medicine R Science Q Shigeo S. Sugano Hiroko Suzuki Eisuke Shimokita Hirofumi Chiba Sumihare Noji Yuriko Osakabe Keishi Osakabe Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| description |
Abstract Mushroom-forming basidiomycetes produce a wide range of metabolites and have great value not only as food but also as an important global natural resource. Here, we demonstrate CRISPR/Cas9-based genome editing in the model species Coprinopsis cinerea. Using a high-throughput reporter assay with cryopreserved protoplasts, we identified a novel promoter, CcDED1 pro , with seven times stronger activity in this assay than the conventional promoter GPD2. To develop highly efficient genome editing using CRISPR/Cas9 in C. cinerea, we used the CcDED1 pro to express Cas9 and a U6-snRNA promoter from C. cinerea to express gRNA. Finally, CRISPR/Cas9-mediated GFP mutagenesis was performed in a stable GFP expression line. Individual genome-edited lines were isolated, and loss of GFP function was detected in hyphae and fruiting body primordia. This novel method of high-throughput CRISPR/Cas9-based genome editing using cryopreserved protoplasts should be a powerful tool in the study of edible mushrooms. |
| format |
article |
| author |
Shigeo S. Sugano Hiroko Suzuki Eisuke Shimokita Hirofumi Chiba Sumihare Noji Yuriko Osakabe Keishi Osakabe |
| author_facet |
Shigeo S. Sugano Hiroko Suzuki Eisuke Shimokita Hirofumi Chiba Sumihare Noji Yuriko Osakabe Keishi Osakabe |
| author_sort |
Shigeo S. Sugano |
| title |
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| title_short |
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| title_full |
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| title_fullStr |
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| title_full_unstemmed |
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system |
| title_sort |
genome editing in the mushroom-forming basidiomycete coprinopsis cinerea, optimized by a high-throughput transformation system |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/42f047196f1f4130a92caee3e7887420 |
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