| RRC ID |
82369
|
| 著者 |
Yamada H, Kato N, Ichikawa M, Mannen K, Kiba T, Osakabe Y, Sakakibara H, Matsui M, Okamoto T.
|
| タイトル |
DNA- and Selectable-Marker-Free Genome-Editing System Using Zygotes from Recalcitrant Maize Inbred B73.
|
| ジャーナル |
Plant Cell Physiol
|
| Abstract |
Genome-editing tools such as the clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9) system have become essential tools for increasing the efficiency and accuracy of plant breeding. Using such genome-editing tools on maize, one of the most important cereal crops of the world, will greatly benefit the agriculture and the mankind. Conventional genome-editing methods typically used for maize involve insertion of a Cas9-guide RNA expression cassette and a selectable marker in the genome DNA; however, using such methods, it is essential to eliminate the inserted DNA cassettes to avoid legislative concerns on gene-modified organisms. Another major hurdle for establishing an efficient and broadly applicable DNA-free genome-editing system for maize is presented by recalcitrant genotypes/cultivars, since cell/tissue culture and its subsequent regeneration into plantlets are crucial for producing transgenic and/or genome-edited maize. In this study, to establish a DNA-free genome-editing system for recalcitrant maize genotypes/cultivars, Cas9-gRNA ribonucleoproteins were directly delivered into zygotes isolated from the pollinated flowers of the maize-B73 cultivar. The zygotes successfully developed and were regenerated into genome-edited plantlets by co-culture with phytosulfokine, a peptide phytohormone. The method developed herein made it possible to obtain DNA- and selectable-marker-free genome-edited recalcitrant maize genotypes/cultivars with high efficiency. This method can advance the molecular breeding of maize and other important cereals, regardless of their recalcitrant characteristics.
|
| 巻・号 |
65(5)
|
| ページ |
729-736
|
| 公開日 |
2024-5-30
|
| DOI |
10.1093/pcp/pcae010
|
| PII |
7591713
|
| PMID |
38288629
|
| MeSH |
CRISPR-Cas Systems*
DNA, Plant / genetics
Gene Editing* / methods
Genome, Plant*
Plant Breeding / methods
Plants, Genetically Modified
RNA, Guide, CRISPR-Cas Systems / genetics
Zea mays* / genetics
Zygote / metabolism
|
| IF |
4.062
|
| リソース情報 |
| シロイヌナズナ / 植物培養細胞・遺伝子 |
rpc00031 |
