J Integr Plant Biol ›› 2023, Vol. 65 ›› Issue (4): 895-899.DOI: 10.1111/jipb.13421

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  • 收稿日期:2022-10-13 接受日期:2022-11-30 出版日期:2023-04-01 发布日期:2023-04-08

An efficient CRISPR/Cas9 platform for targeted genome editing in rose (Rosa hybrida)

Chengpeng Wang1#, Yang Li1, Na Wang1, Qin Yu1, Yonghong Li2, Junping Gao1, Xiaofeng Zhou1* and Nan Ma1*   

  1. 1. Beijing Key Laboratory of Development and Quality Control of Ornamental Crops, Department of Ornamental Horticulture, China Agricultural University, Beijing 100193, China;
    2. School of Applied Chemistry and Biotechnology, Shenzhen Polytechnic, Shenzhen 518038, China;
    #Current Address: Key Laboratory of East China Urban Agriculture, Ministry of Agriculture and Rural Affairs, Institute of Leisure Agriculture, Shandong Academy of Agricultural Sciences, Jinan 250100, China
    *Correspondences: Nan Ma (ma_nan@cau.edu.cn, Dr. Ma is responsible for the distribution of the material associated with this article); Xiaofeng Zhou (zhouxiaofeng@cau.edu.cn)
  • Received:2022-10-13 Accepted:2022-11-30 Online:2023-04-01 Published:2023-04-08

Abstract: The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐related nuclease 9 (Cas9) system enables precise, simple editing of genes in many animals and plants. However, this system has not been applied to rose (Rosa hybrida) due to the genomic complexity and lack of an efficient transformation technology for this plant. Here, we established a platform for screening single‐guide RNAs (sgRNAs) with high editing efficiency for CRISPR/Cas9‐mediated gene editing in rose using suspension cells. We used the Arabidopsis thaliana U6‐29 promoter, which showed high activity for driving sgRNA expression, to modify the CRISPR/Cas9 system. We used our highly efficient optimized CRISPR/Cas9 system to successfully edit RhEIN2, encoding an indispensable component of the ethylene signaling pathway, resulting in ethylene insensitivity in rose. Our optimized CRISPR/Cas9 system provides a powerful toolbox for functional genomics, molecular breeding, and synthetic biology in rose.

Key words: CRISPR/CAS9, gene editing, RhEIN2, rose

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