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| CRISPR-Cas9-mediated genome editing in apple and grapevine | |
| Osakabe, Yuriko; Liang, Zhenchang1; Ren, Chong1; Nishitani, Chikako2; Osakabe, Keishi; Wada, Masato3; Komori, Sadao4; Malnoy, Mickael5; Velasco, Riccardo5,9; Poli, Michele; Jung, Min-Hee; Koo, Ok-Jae; Viola, Roberto8; Kanchiswamy, Chidananda Nagamangala5,6 | |
| 2018 | |
| 发表期刊 | NATURE PROTOCOLS
 |
| ISSN | 1754-2189 |
| 卷号 | 13期号:12页码:2844-2863 |
| 摘要 | The CRISPR-Cas9 genome-editing tool and the availability of whole-genome sequences from plant species have revolutionized our ability to introduce targeted mutations into important crop plants, both to explore genetic changes and to introduce new functionalities. Here, we describe protocols adapting the CRISPR-Cas9 system to apple and grapevine plants, using both plasmid-mediated genome editing and the direct delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) to achieve efficient DNA-free targeted mutations in apple and grapevine protoplasts. We provide a stepwise protocol for the design and transfer of CRISPR-Cas9 components to apple and grapevine protoplasts, followed by verification of highly efficient targeted mutagenesis, and regeneration of plants following the plasmid-mediated delivery of components. Our plasmid-mediated procedure and the direct delivery of CRISPR-Cas9 RNPs can both be utilized to modulate traits of interest with high accuracy and efficiency in apple and grapevine, and could be extended to other crop species. The complete protocol employing the direct delivery of CRISPR-Cas9 RNPs takes as little as 2-3 weeks, whereas the plasmid-mediated procedure takes >3 months to regenerate plants and study the mutations. |
| 学科领域 | Biochemical Research Methods |
| DOI | 10.1038/s41596-018-0067-9 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS关键词 | LEAF MESOPHYLL PROTOPLASTS ; SOMATIC EMBRYOGENESIS ; PLANT-REGENERATION ; GENETIC-TRANSFORMATION ; TARGETED MUTAGENESIS ; HUMAN-CELLS ; DNA ; CRISPR/CAS9 ; DELIVERY ; SYSTEM |
| WOS记录号 | WOS:000451343400007 |
| 出版者 | NATURE PORTFOLIO |
| 文献子类 | Article |
| 出版地 | BERLIN |
| EISSN | 1750-2799 |
| 资助机构 | Council for Science, Technology and Innovation (CSTI) ; Cross-ministerial Strategic Innovation Promotion Program (SIP) ; 'Technologies for creating next-generation agriculture, forestry and fisheries' program (funding agency: Bio-oriented Technology Research Advancement Institution, NARO) in Japan ; Program on Open Innovation Platform with Enterprises, Research Institute and Academia (OPERA) in Japan ; Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, and Forestry (IPET) through the Agri-Bio Industry Technology Development Program - Ministry of Agriculture, Food and Rural Affairs (MAFRA) [116088-3] |
| 作者邮箱 | osakabe.yuriko@tokushima-u.ac.jp ; chida@plantedit.com |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.ibcas.ac.cn/handle/2S10CLM1/20584 |
| 专题 | 中科院北方资源植物重点实验室 |
| 作者单位 | 1.Tokushima Univ, Fac Biosci & Bioind, Tokushima, Tokushima, Japan 2.Chinese Acad Sci, Beijing Key Lab Grape Sci & Enol, Inst Bot, Key Lab Plant Resources, Beijing, Peoples R China 3.NARO Inst Fruit Tree & Tea Sci, Div Fruit Breeding & Genet, Tsukuba, Ibaraki, Japan 4.NARO Inst Fruit Tree & Tea Sci, Div Apple Res, Morioka, Iwate, Japan 5.Iwate Univ, Fac Agr, Morioka, Iwate, Japan 6.FEM, Genom & Biol Fruit Crop Dept, Res & Innovat Ctr, San Michele All Adige, TN, Italy 7.PLANTeDIT Pvt Ltd, Cork, Ireland 8.ToolGen, Seoul, South Korea 9.Italian Inst Technol, Genoa, Italy 10.CREA, Res Ctr Viticulture & Enol, Conegliano, TV, Italy |
| 推荐引用方式 GB/T 7714 | Osakabe, Yuriko,Liang, Zhenchang,Ren, Chong,et al. CRISPR-Cas9-mediated genome editing in apple and grapevine[J]. NATURE PROTOCOLS,2018,13(12):2844-2863. |
| APA | Osakabe, Yuriko.,Liang, Zhenchang.,Ren, Chong.,Nishitani, Chikako.,Osakabe, Keishi.,...&Kanchiswamy, Chidananda Nagamangala.(2018).CRISPR-Cas9-mediated genome editing in apple and grapevine.NATURE PROTOCOLS,13(12),2844-2863. |
| MLA | Osakabe, Yuriko,et al."CRISPR-Cas9-mediated genome editing in apple and grapevine".NATURE PROTOCOLS 13.12(2018):2844-2863. |
| 条目包含的文件 | ||||||
| 文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
| 30390050.pdf（1650KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | 浏览 请求全文 | |
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