茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建

被引：10

作者：

唐雨薇 ^{[1
,2
]}

刘丽萍 ^{[1
,2
]}

王若娴 ^{[1
]}

陈宇宏 ^{[1
]}

刘仲华 ^{[1
,3
,2
]}

刘硕谦 ^{[1
,3
,2
]}

机构：

[1] 湖南农业大学园艺园林学院

[2] 教育部茶学重点实验室

[3] 国家植物功能成分利用工程技术研究中心

来源：

茶叶科学 | 2016年 / 36卷 / 04期

关键词：

茶树; 基因组编辑技术; 咖啡碱合成酶; CRISPR/Cas9技术;

D O I：

10.13305/j.cnki.jts.2016.04.010

中图分类号：

S571.1 [茶]; Q943.2 [植物基因工程];

学科分类号：

0902 ; 090203 ; 071007 ; 090102 ;

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。

引用

页码：414 / 426

页数：13

共 10 条

[1]

Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System[J]. Zhen Liang,Kang Zhang,Kunling Chen,Caixia Gao.Journal of Genetics and Genomics. 2014(02)

[2]

CRISPR/Cas9-mediated genome editing and gene replacement in plants: Transitioning from lab to field[J] . Scott M. Schaeffer,Paul A. Nakata.Plant Science . 2015

[3] Genome-Wide Identification of Genes Probably Relevant to the Uniqueness of Tea Plant (Camellia sinensis) and Its Cultivars [J].

Yan Wei ;

Wang Jing ;

Zhou Youxiang ;

Zhao Mingming ;

Gong Yan ;

Ding Hua ;

Peng Lijun ;

Hu Dingjin .

INTERNATIONAL JOURNAL OF GENOMICS, 2015, 2015

[4]

A Robust CRISPR/Cas9 System for Convenient, High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants[J] . Xingliang Ma,Qunyu Zhang,Qinlong Zhu,Wei Liu,Yan Chen,Rong Qiu,Bin Wang,Zhongfang Yang,Heying Li,Yuru Lin,Yongyao Xie,Rongxin Shen,Shuifu Chen,Zhi Wang,Yuanling Chen,Jingxin Guo,Letian Chen,Xiucai Zhao,Zhicheng Dong,Yao-Guang Liu.Molecular Plant . 2015 (8)

[5]

Editing plant genomes with CRISPR/Cas9[J] . Khaoula Belhaj,Angela Chaparro-Garcia,Sophien Kamoun,Nicola J Patron,Vladimir Nekrasov.Current Opinion in Biotechnology . 2015

[6]

RNA-Guided Genome Editing in Plants Using a CRISPR–Cas System[J] . Kabin Xie,Yinong Yang.Molecular Plant . 2013 (6)

[7] CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes [J].

Gilbert, Luke A. ;

Larson, Matthew H. ;

Morsut, Leonardo ;

Liu, Zairan ;

Brar, Gloria A. ;

Torres, Sandra E. ;

Stern-Ginossar, Noam ;

Brandman, Onn ;

Whitehead, Evan H. ;

Doudna, Jennifer A. ;

Lim, Wendell A. ;

Weissman, Jonathan S. ;

Qi, Lei S. .

CELL, 2013, 154 (02) :442-451

[8] One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering [J].

Wang, Haoyi ;

Yang, Hui ;

Shivalila, Chikdu S. ;

Dawlaty, Meelad M. ;

Cheng, Albert W. ;

Zhang, Feng ;

Jaenisch, Rudolf .

CELL, 2013, 153 (04) :910-918

[9]

Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice[J] . Wenzhi Jiang,Huanbin Zhou,Honghao Bi,Michael Fromm,Bing Yang,Donald P. Weeks.Nucleic Acids Research . 2013 (20)

[10]

The Structure of the CRISPR-Associated Protein Csa3 Provides Insight into the Regulation of the CRISPR/Cas System[J] . Nathanael G. Lintner,Kenneth A. Frankel,Susan E. Tsutakawa,Donald L. Alsbury,Valérie Copié,Mark J. Young,John A. Tainer,C. Martin Lawrence.Journal of Molecular Biology . 2010 (4)

← 1 →