CRISPR-Cas9-mediated genome editing in apple and grapevine

doi:10.1038/s41596-018-0067-9

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	CRISPR-Cas9-mediated genome editing in apple and grapevine
	Osakabe, Yuriko ; Liang, Zhenchang 9; Ren, Chong 9; Nishitani, Chikako 8; Osakabe, Keishi ; Wada, Masato 7; Komori, Sadao 6; Malnoy, Mickael 5; Velasco, Riccardo 4,5; Poli, Michele
刊名	NATURE PROTOCOLS
	2018
卷号	13 期号:12 页码:2844-2863
ISSN号	1754-2189
DOI	10.1038/s41596-018-0067-9
文献子类	Article
英文摘要	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.
学科主题	Biochemical Research Methods
电子版国际标准刊号	1750-2799
出版地	BERLIN
WOS关键词	LEAF MESOPHYLL PROTOPLASTS ; SOMATIC EMBRYOGENESIS ; PLANT-REGENERATION ; GENETIC-TRANSFORMATION ; TARGETED MUTAGENESIS ; HUMAN-CELLS ; DNA ; CRISPR/CAS9 ; DELIVERY ; SYSTEM
语种	英语
出版者	NATURE PORTFOLIO
WOS记录号	WOS:000451343400007
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]
内容类型	期刊论文
源URL	[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.CREA, Res Ctr Viticulture & Enol, Conegliano, TV, Italy 2.PLANTeDIT Pvt Ltd, Cork, Ireland 3.ToolGen, Seoul, South Korea 4.Italian Inst Technol, Genoa, Italy 5.Iwate Univ, Fac Agr, Morioka, Iwate, Japan 6.NARO Inst Fruit Tree & Tea Sci, Div Apple Res, Morioka, Iwate, Japan 7.NARO Inst Fruit Tree & Tea Sci, Div Fruit Breeding & Genet, Tsukuba, Ibaraki, Japan 8.Chinese Acad Sci, Beijing Key Lab Grape Sci & Enol, Inst Bot, Key Lab Plant Resources, Beijing, Peoples R China 9.Tokushima Univ, Fac Biosci & Bioind, Tokushima, Tokushima, Japan 10.FEM, Genom & Biol Fruit Crop Dept, Res & Innovat Ctr, San Michele All Adige, TN, 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.