A detailed procedure for CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana

查看全文 作　　者：Wenshan [1,2,3]Liu;Xiaohong [2]Zhu;Mingguang [4]Lei;Qingyou [3]Xia;Jose Ramon [5]Botella;Jian-Kang [2,4]Zhu;Yanfei [2]Mao

机构地区：[1]School of Life Sciences, Chongqing University;[2]Shanghai Center for Plant Stress Biology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences;[3]State Key Laboratory of Silkworm Genome Biology, Southwest University;[4]Department of Horticulture and Landscape Architecture, Purdue University;[5]School of Agriculture and Food Sciences, University of Queensland

出　　处：《Science Bulletin》索引2015年第60卷第15期,共16页

基　　金：supported by the Chinese Academy of Sciences and China Scholarship Council(201206050103) 摘　　要：The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesis and transgenic methods to study gene function and improve crop traits. CRISPR/Cas facilitates targeted gene editing through RNA-guided DNA cleavage followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. Here we describe a detailed procedure for our previously developed and highly efficient CRISPR/Cas9 method that allows the generation of heritable-targeted gene mutations andcorrections in Arabidopsis. This protocol describes the strategies and steps for the selection of targets, design of single-guide RNA(sg RNA), vector construction and analysis of transgenic lines. We also offer a method to target two loci simultaneously using vectors containing two different sg RNAs. The principles described in this protocol can be applied to other plant species to generate stably inherited DNA modifications. 关键词：RNA编辑基因组拟南芥转基因载体 DNA裂解介导程序协议描述

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A detailed procedure for CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana

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