| Autor: |
Shigeo S Sugano, Ryuichi Nishihama, Makoto Shirakawa, Junpei Takagi, Yoriko Matsuda, Sakiko Ishida, Tomoo Shimada, Ikuko Hara-Nishimura, Keishi Osakabe, Takayuki Kohchi |
| Jazyk: |
angličtina |
| Rok vydání: |
2018 |
| Předmět: |
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| Zdroj: |
PLoS ONE, Vol 13, Iss 10, p e0205117 (2018) |
| Druh dokumentu: |
article |
| ISSN: |
1932-6203 |
| DOI: |
10.1371/journal.pone.0205117 |
| Popis: |
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. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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