Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha.
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 w...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC6209168?pdf=render |
| _version_ | 1818059557436391424 |
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| author | Shigeo S Sugano Ryuichi Nishihama Makoto Shirakawa Junpei Takagi Yoriko Matsuda Sakiko Ishida Tomoo Shimada Ikuko Hara-Nishimura Keishi Osakabe Takayuki Kohchi |
| author_facet | Shigeo S Sugano Ryuichi Nishihama Makoto Shirakawa Junpei Takagi Yoriko Matsuda Sakiko Ishida Tomoo Shimada Ikuko Hara-Nishimura Keishi Osakabe Takayuki Kohchi |
| author_sort | Shigeo S Sugano |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T13:18:25Z |
| format | Article |
| id | doaj.art-9f2622d2a4a6491aa3b9e0a4b4656000 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T13:18:25Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9f2622d2a4a6491aa3b9e0a4b46560002022-12-22T01:47:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011310e020511710.1371/journal.pone.0205117Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha.Shigeo S SuganoRyuichi NishihamaMakoto ShirakawaJunpei TakagiYoriko MatsudaSakiko IshidaTomoo ShimadaIkuko Hara-NishimuraKeishi OsakabeTakayuki KohchiMarchantia 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.http://europepmc.org/articles/PMC6209168?pdf=render |
| spellingShingle | Shigeo S Sugano Ryuichi Nishihama Makoto Shirakawa Junpei Takagi Yoriko Matsuda Sakiko Ishida Tomoo Shimada Ikuko Hara-Nishimura Keishi Osakabe Takayuki Kohchi Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLoS ONE |
| title | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. |
| title_full | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. |
| title_fullStr | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. |
| title_full_unstemmed | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. |
| title_short | Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. |
| title_sort | efficient crispr cas9 based genome editing and its application to conditional genetic analysis in marchantia polymorpha |
| url | http://europepmc.org/articles/PMC6209168?pdf=render |
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