A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants

扫码查看

原文链接

NETL
NSTL
万方数据

外文摘要：The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombi-nation to assemble large fragments and by changing the structure of the amplicons used to assemble sgRNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single bin-ary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sgRNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex gen-ome editing system advances synthetic biology and plant genetic engineering.

外文关键词：

CRISPR/Cas9Multiplex genome editingAssembly systemPlant

作者：

Suting Wu、Htin Kyaw、Zhijun Tong、Yirong Yang、Zhiwei Wang、Liying Zhang、Lihua Deng、Zhiguo Zhang、Bingguang Xiao、William Paul Quick、Tiegang Lu、Guoying Xiao、Guannan Qin、Xue'an Cui

展开 >

作者单位：

Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China

Key Laboratory of Tobacco Biotechnological Breeding,National Tobacco Genetic Engineering Research Center,Yunnan Academy of Tobacco Agricultural Sciences,Kunming 650021,Yunnan,China

Key Laboratory of Agro-ecological Processes in Subtropical Region,Institute of Subtropical Agriculture,Chinese Academy of Sciences,Changsha 410125,Hunan,China

School of Biosciences,The University of Sheffield,Firth Court,Western Bank,Sheffield S10 2TN,UK

Key Laboratory of Genetics,Breeding and Multiple Utilization of Crops,Ministry of Education,College of Agriculture,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China

Key Laboratory of Biological Breeding for Fujian and Taiwan Crops,Ministry of Agriculture and Rural Affairs,College of Agriculture,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China

展开 >

基金：

国家自然科学基金国家自然科学基金国家重点研发计划Hunan Seed Industry Innovation ProjectYunnan Tobacco Company Project

项目编号：

32001532318604112022YFF10000202021NK10122020530000241009

出版年：

2024

DOI：

10.1016/j.cj.2024.01.010

作物学报(英文版)

CSTPCD

ISSN：

年,卷(期)：2024.12(2)

参考文献量62