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タイトル: | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha |
著者: | Sugano, Shigeo S. Nishihama, Ryuichi https://orcid.org/0000-0002-7032-732X (unconfirmed) Shirakawa, Makoto Takagi, Junpei Matsuda, Yoriko Ishida, Sakiko Shimada, Tomoo Hara-Nishimura, Ikuko Osakabe, Keishi Kohchi, Takayuki https://orcid.org/0000-0002-9712-4872 (unconfirmed) |
著者名の別形: | 西浜, 竜一 松田, 頼子 石田, 咲子 西村, いくこ 河内, 孝之 |
発行日: | 31-Oct-2018 |
出版者: | Public Library of Science (PLoS) |
誌名: | PLOS ONE |
巻: | 13 |
号: | 10 |
論文番号: | e0205117 |
抄録: | Marchantia polymorpha is one of the model species of basal land plants. Although CRISPR/Cas9-based genome editing has already been demonstrated for this plant, the efficiency was too low to apply to functional analysis. In this study, we show the establishment of CRISPR/Cas9 genome editing vectors with high efficiency for both construction and genome editing. Codon optimization of Cas9 to Arabidopsis achieved over 70% genome editing efficiency at two loci tested. Systematic assessment revealed that guide sequences of 17 nt or shorter dramatically decreased this efficiency. We also demonstrated that a combinatorial use of this system and a floxed complementation construct enabled conditional analysis of a nearly essential gene. This study reports that simple, rapid, and efficient genome editing is feasible with the series of developed vectors. |
著作権等: | © 2018 Sugano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
URI: | http://hdl.handle.net/2433/234941 |
DOI(出版社版): | 10.1371/journal.pone.0205117 |
PubMed ID: | 30379827 |
出現コレクション: | 学術雑誌掲載論文等 |
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