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

TANG Yuwei; LIU Liping; WANG Ruoxian; CHEN Yuhong; LIU Zhonghua; LIU Shuoqian

doi:10.13305/j.cnki.jts.2016.04.010

Journal of Tea Science >

2016 , Vol. 36 >Issue 4: 414 - 426

DOI: https://doi.org/10.13305/j.cnki.jts.2016.04.010

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

TANG Yuwei ,
LIU Liping ,
WANG Ruoxian ,
CHEN Yuhong ,
LIU Zhonghua ,
LIU Shuoqian

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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 date: 2016-04-25

Online published: 2019-08-26

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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.

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

Cite this article

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 , 36(4) : 414 -426 . DOI: 10.13305/j.cnki.jts.2016.04.010

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