Repositorium
Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha
350
Book / 2018
Sugano, Shigeo S.; Nishihama, Ryuichi; Shirakawa, Makoto; Takagi, Junpei; Matsuda, Yoriko; Ishida, Sakiko; Shimada, Tomoo; Hara-Nishimura, Ikuko; Osakabe, Keishi; Kohchi, Takayuki
Marchantia polymorpha is one of the model species of basal land plants. Although CRISPR/Cas-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.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
819 |
Kohchi; Takayuki Japan |
Marchantia polymorpha |
CRISPR/Cas9 ARF1 |
NAA resistance SDN1 |
Basic research Basic research |
820 |
Kohchi; Takayuki Japan |
Marchantia polymorpha |
CRISPR/Cas9 NOP1 |
Air chamber formation SDN1 |
Basic research Basic research |
821 |
Kohchi; Takayuki Japan |
Marchantia polymorpha |
CRISPR/Cas9 MPK1 |
Regulates growth and development SDN1 |
Basic research Basic research |