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

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Journal of Tea Science ›› 2016, Vol. 36 ›› Issue (4) : 414-426. DOI: 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^1,3, LIU Liping^1,3, WANG Ruoxian¹, CHEN Yuhong¹, LIU Zhonghua^1,2,3, LIU Shuoqian^1,2,3,*

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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.) / CRISPR/Cas9 technique / genome editing technology / tea caffeine synthase

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

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