Repositorium
Journal Article / 2016
Nomura, Toshihisa; Sakurai, Tetsuya; Osakabe, Yuriko; Osakabe, Keishi; Sakakibara, Hitoshi
Targeted genome modification by RNA-guided nucleases derived from the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has seen rapid development in many organisms, including several plant species. In the present study, we succeeded in introducing the CRISPR/Cas9 system into the non-model organism Scopelophila cataractae, a moss that exhibits heavy metal tolerance, and the model organism Physcomitrella patens Utilizing the process by which moss plants regenerate from protoplasts, we conducted targeted mutagenesis by expression of single-chain guide RNA (sgRNA) and Cas9 in protoplasts. Using this method, the acquisition rate of strains exhibiting phenotypic changes associated with the target genes was approximately 45-69%, and strains with phenotypic changes exhibited various insertion and deletion mutations. In addition, we report that our method is capable of multiplex targeted mutagenesis (two independent genes) and also permits the efficient introduction of large deletions (∼3 kbp). These results demonstrate that the CRISPR/Cas9 system can be used to accelerate investigations of bryology and land plant evolution.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
338 |
Sakakibara, Hitoshi. Toshihisa Nomura Japan |
Physcomitrella patens |
CRISPR/Cas9 FtsZ2-1 |
responsible for chloroplast division SDN1 |
Basic research Basic research |
339 |
Sakakibara, Hitoshi. Toshihisa Nomura Japan |
Physcomitrella patens |
CRISPR/Cas9 TON |
small gametophores with short and thickened leaves SDN1 |
Basic research Basic research |
340 |
Sakakibara, Hitoshi. Toshihisa Nomura Japan |
Scopelophila cataractae |
CRISPR/Cas9 FtsZ2-1 |
responsible for chloroplast division SDN1 |
Basic research Basic research |