Construct design for CRISPR/Cas-based genome editing in plants

被引:84
作者
Hassan, Md Mahmudul [1 ,2 ,3 ]
Zhang, Yingxiao [4 ]
Yuan, Guoliang [1 ,2 ]
De, Kuntal [1 ]
Chen, Jin-Gui [1 ,2 ]
Muchero, Wellington [1 ,2 ]
Tuskan, Gerald A. [1 ,2 ]
Qi, Yiping [4 ,5 ]
Yang, Xiaohan [1 ,2 ]
机构
[1] Oak Ridge Natl Lab, Biosci Div, Oak Ridge, TN 37831 USA
[2] Oak Ridge Natl Lab, Ctr Bioenergy Innovat, Oak Ridge, TN 37831 USA
[3] Patuakhali Sci & Technol Univ, Dept Genet & Plant Breeding, Dumki 8602, Patuakhali, Bangladesh
[4] Univ Maryland, Dept Plant Sci & Landscape Architecture, College Pk, MD 20742 USA
[5] Univ Maryland, Inst Biosci & Biotechnol Res, Rockville, MD 20850 USA
基金
美国国家科学基金会;
关键词
HIGHLY EFFICIENT; GUIDE RNA; MULTIPLEX; DNA; MUTAGENESIS; RICE; ENDONUCLEASE; ARABIDOPSIS; EXPRESSION; INDUCTION;
D O I
10.1016/j.tplants.2021.06.015
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
CRISPR construct design is a key step in the practice of genome editing, which includes identification of appropriate Cas proteins, design and selection of guide RNAs (gRNAs), and selection of regulatory elements to express gRNAs and Cas proteins. Here, we review the choices of CRISPR-based genome editors suited for different needs in plant genome editing applications. We consider the technical aspects of gRNA design and the associated computational tools. We also discuss strategies for the design of multiplex CRISPR constructs for highthroughput manipulation of complex biological processes or polygenic traits. We provide recommendations for different elements of CRISPR constructs and discuss the remaining challenges of CRISPR construct optimization in plant genome editing.
引用
收藏
页码:1133 / 1152
页数:20
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