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Journal of Tea Science ›› 2016, Vol. 36 ›› Issue (4): 414-426.doi: 10.13305/j.cnki.jts.2016.04.010

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Development of a CRISPR/Cas9 Constructed for Genome Editing of Caffeine Synthase in Camellia sinensis

TANG Yuwei1,3, LIU Liping1,3, WANG Ruoxian1, CHEN Yuhong1, LIU Zhonghua1,2,3, LIU Shuoqian1,2,3,*   

  1. 1. College of Horticulture and Hardening, Hunan Agricultural University, Changsha 410128, China;
    2. National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, Changsha 410128, China;
    3. Key Lab of Tea Science, Ministry of Education, Changsha 410128, China
  • Received:2016-04-25 Published:2019-08-26

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.

Key words: Camellia sinensis (L.), genome editing technology, tea caffeine synthase, CRISPR/Cas9 technique

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