Articles
Efficient genome editing in carnation (Dianthus caryophyllus) using the CRISPR/Cas9 system
Article number
1383_23
Pages
193 – 200
Language
English
Abstract
Carnation (Dianthus caryophyllus) is one of the most popular and widely used ornamental flowers worldwide.
Although the genome sequencing and the mechanism of flower color formation and petal senescence have been reported, so far efficient CRISPR-Cas genome editing toolkits in carnation have not been established.
In this study, we identified a DcU6 promoter, and also established a fast and effective carnation protoplast screening system using NanoLUC as the reporter to screen the high-efficiency elements in CRISPR/Cas9 system.
The results demonstrated that the DcU6 promoter exhibited a more effective role than the AtU6 promoter in editing efficiency in carnation protoplast.
Besides, we found that the SlEF1α promoter from tomato could significantly increase the editing efficiency by driving the expression of AtCas9. DcPDS was chosen as a target to further identify the editing efficiency using the CRISPR/Cas9 system containing the DcU6-1 promoter, SlEF1α promoter and AtCas9. Our study provides an optimization of the CRISPR/Cas9 system in carnation.
The application of gene editing technologies will be used for the improvement, especially for disease resistance, abiotic stress tolerance, herbicide tolerance, shelf life, and yield in flower plants.
Although the genome sequencing and the mechanism of flower color formation and petal senescence have been reported, so far efficient CRISPR-Cas genome editing toolkits in carnation have not been established.
In this study, we identified a DcU6 promoter, and also established a fast and effective carnation protoplast screening system using NanoLUC as the reporter to screen the high-efficiency elements in CRISPR/Cas9 system.
The results demonstrated that the DcU6 promoter exhibited a more effective role than the AtU6 promoter in editing efficiency in carnation protoplast.
Besides, we found that the SlEF1α promoter from tomato could significantly increase the editing efficiency by driving the expression of AtCas9. DcPDS was chosen as a target to further identify the editing efficiency using the CRISPR/Cas9 system containing the DcU6-1 promoter, SlEF1α promoter and AtCas9. Our study provides an optimization of the CRISPR/Cas9 system in carnation.
The application of gene editing technologies will be used for the improvement, especially for disease resistance, abiotic stress tolerance, herbicide tolerance, shelf life, and yield in flower plants.
Authors
Xueqing Fu, Kexuan Tang
Keywords
gene editing, CRISPR/Cas9, Dianthus caryophyllus, U6 promoter
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