| Item type |
学術雑誌論文 / Journal Article(1) |
| 公開日 |
2024-11-01 |
| タイトル |
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|
タイトル |
Development of a gene-targeting system using CRISPR/Cas9 and utilization of pyrG as a novel selectable marker in Lentinula edodes |
|
言語 |
en |
| 言語 |
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|
言語 |
eng |
| 資源タイプ |
|
|
資源 |
http://purl.org/coar/resource_type/c_6501 |
|
タイプ |
journal article |
| アクセス権 |
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|
アクセス権 |
open access |
|
アクセス権URI |
http://purl.org/coar/access_right/c_abf2 |
| 著者 |
Ayane Kamiya
Hiroki Ueshima
Shota Nishida
Yoichi Honda
Hisatoshi Kamitsuji
Toshitsugu Sato
Haruto Miyamoto
Takuya Sumita
Kosuke Izumitsu
Toshikazu Irie
|
| 抄録 |
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内容記述タイプ |
Abstract |
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内容記述 |
First, we attempted to recombine the Shiitake (Lentinula edodes) pyrG (ura3) gene homologously by introducing a donor vector containing a carboxin resistance gene (lecbxR) flanked by homologous sequences of pyrG into protoplasts of the fungus. However, all the carboxin-resistant transformants only contained ectopic insertions of the exogenous gene and no homologous insertions. Agaricomycetes are generally known for their low efficiency of homologous recombination, and a similar result was shown for L. edodes. We then co-introduced a Cas9 plasmid vector containing a CRISPR/Cas9 expression cassette targeting pyrG and donor plasmid vector. As a result, ∆pyrG strains containing the expected homologous recombination were obtained. However, only two of the seven ∆pyrG strains had the Cas9 sequence; the others did not. Our results suggest that genome editing occurred via the transient expression of the CRISPR/Cas9 cassette in the Cas9 plasmid vector introduced into the fungal cell. Transforming pyrG into a ∆pyrG strain (strain I8) resulted in prototrophic strains with an efficiency of 6.5 strains/experiment. |
|
言語 |
en |
| 書誌情報 |
en : FEMS Microbiology Letters
巻 370,
p. 1-6,
発行日 2023-05
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| ISSN |
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収録物識別子タイプ |
EISSN |
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収録物識別子 |
1574-6968 |
| DOI |
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|
識別子タイプ |
DOI |
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関連識別子 |
10.1093/femsle/fnad042 |
| 権利 |
|
|
言語 |
en |
|
権利情報 |
c2023 Oxford University Press This is a pre-copyedited, author-produced version of an article accepted for publication in FEMS Microbiology Letters following peer review. The version of record Ayane Kamiya, Hiroki Ueshima, Shota Nishida, Yoichi Honda, Hisatoshi Kamitsuji, Toshitsugu Sato, Haruto Miyamoto, Takuya Sumita, Kosuke Izumitsu, Toshikazu Irie, Development of a gene-targeting system using CRISPR/Cas9 and utilization of pyrG as a novel selectable marker in Lentinula edodes, FEMS Microbiology Letters, Volume 370, 2023, fnad042, is available online at: https://doi.org/10.1093/femsle/fnad042 |
| 出版者 |
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|
出版者 |
Oxford University Press |
|
言語 |
en |
| 著者版フラグ |
|
|
言語 |
en |
|
値 |
author |
| 出版タイプ |
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|
出版タイプ |
AM |
|
出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
article.pdf (2.1 MB)
