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タイトル: | Efficient genome editing with CRISPR/Cas9 in Pleurotus ostreatus |
著者: | Boontawon, Tatpong Nakazawa, Takehito https://orcid.org/0000-0002-9654-2124 (unconfirmed) Inoue, Chikako Osakabe, Keishi Kawauchi, Moriyuki https://orcid.org/0000-0001-7673-5896 (unconfirmed) Sakamoto, Masahiro Honda, Yoichi https://orcid.org/0000-0002-7218-7367 (unconfirmed) |
著者名の別形: | 中沢, 威人 井上, 智香子 刑部, 敬史 河内, 護之 坂本, 正弘 本田, 与一 |
キーワード: | Agaricomycete Mushroom fcy1 pyrG Genome editing |
発行日: | 20-Feb-2021 |
出版者: | Springer Nature |
誌名: | AMB Express |
巻: | 11 |
論文番号: | 30 |
抄録: | Pleurotus ostreatus is one of the most commercially produced edible mushrooms worldwide. Improved cultivated strains with more useful traits have been obtained using classical breeding, which is laborious and time-consuming. Here, we attempted efficient gene mutagenesis using plasmid-based CRISPR/Cas9 as the first step for non-genetically modified (non-GM) P. ostreatus generation. Plasmids harboring expression cassettes of Cas9 and different single guide RNAs targeting fcy1 and pyrG were individually transferred into fungal protoplasts of the PC9 strain, which generated some strains exhibiting resistance to 5-fluorocytosine and 5-fluoroorotic acid, respectively. Genomic PCR followed by sequencing revealed small insertions/deletions or insertion of a fragment from the plasmid at the target site in some of the drug-resistant strains. The results demonstrated efficient CRISPR/Cas9-assisted genome editing in P. ostreatus, which could contribute to the molecular breeding of non-GM cultivated strains in the future. Furthermore, a mutation in fcy1 via homology-directed repair using this CRISPR/Cas9 system was also efficiently introduced, which could be applied not only for precise gene disruption, but also for insertions leading to heterologous gene expression in this fungus. |
著作権等: | © The Author(s) 2021. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
URI: | http://hdl.handle.net/2433/261733 |
DOI(出版社版): | 10.1186/s13568-021-01193-w |
PubMed ID: | 33609205 |
出現コレクション: | 学術雑誌掲載論文等 |
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