| 茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建 |
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| 引用本文： | 唐雨薇,刘丽萍,王若娴,陈宇宏,刘仲华,刘硕谦.茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建[J].茶叶科学,2016(4):414-426. |
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| 作者姓名： | 唐雨薇 刘丽萍 王若娴 陈宇宏 刘仲华 刘硕谦 |
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| 作者单位： | 1. 湖南农业大学园艺园林学院，湖南 长沙 410128; 教育部茶学重点实验室，湖南 长沙 410128;2. 湖南农业大学园艺园林学院,湖南 长沙,410128;3. 湖南农业大学园艺园林学院，湖南 长沙 410128; 国家植物功能成分利用工程技术研究中心，湖南 长沙 410128; 教育部茶学重点实验室，湖南 长沙 410128 |
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| 摘 要： | CRISPR/Cas9技术是一门新兴的基因组定点编辑技术,具有操作简单、高效的优点,可轻松实现对目标基因的敲除、替换和定点突变等操作。该技术刚诞生,就受到了全球生命科学领域研究者的关注,不到3年的时间就已经成功应用于多种动、植物当中。然而CRISPR/Cas9技术在茶树中的应用面临载体构建问题,本文以茶树咖啡碱合成酶为例,联合采用常规PCR、Overlapping PCR和Golden Gate Cloning技术,构建了包含茶树咖啡碱合成酶双靶点的CRISPR/Cas9基因编辑载体,为CRISPR/Cas9介导的基因组编辑技术在茶树中的应用奠定了坚实基础。
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| 关 键 词： | 茶树 基因组编辑技术 咖啡碱合成酶 CRISPR/Cas9 技术 |
Development of a CRISPR/Cas9 Constructed for Genome Editing of Caffeine Synthase in Camellia sinensis |
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| TANG Yuwei,LIU Liping,WANG Ruoxian,CHEN Yuhong,LIU Zhonghua,LIU Shuoqian.Development of a CRISPR/Cas9 Constructed for Genome Editing of Caffeine Synthase in Camellia sinensis[J].Journal of Tea Science,2016(4):414-426. |
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| Authors: | TANG Yuwei LIU Liping WANG Ruoxian CHEN Yuhong LIU Zhonghua LIU Shuoqian |
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| Abstract: | CRISPR/Cas9 technology (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) is a novel and powerful approach for targeted genome editing, such as targeted gene knock out or site-directed mutagenesis in a simple and easy way. Since its establishment, the CRISPR/Cas9 technique has been successfully applied in many eukaryotic organisms, including more than 10 plant species. However, it has not been available for genome editing of tea plant Camellia sinensis (L.) O. Kuntze] due to the difficulty in constructing CRISPR/Cas9 expression vector. The present work developed an efficient method to construct a CRISPR/Cas9 expression vector for genome editing a tea caffeine synthase (TCS) by using general PCR, overlapping PCR and golden gate cloning technology. The present work would promote the application of CRISPR/Cas9 technology in genomic modification in tea plants. |
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| Keywords: | Camellia sinensis (L ) genome editing technology tea caffeine synthase CRISPR/Cas9 technique |
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