An efficient CRISPR/Cas9 platform for targeted genome editing in rose (Rosa hybrida) |
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Authors: | Chengpeng Wang Yang Li Na Wang Qin Yu Yonghong Li Junping Gao Xiaofeng Zhou Nan Ma |
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Affiliation: | 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 |
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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. |
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Keywords: | CRISPR/CAS9 gene editing RhEIN2 rose |
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