A CRISPR/dCas9 toolkit for functional analysis of maize genes

被引：22

作者：

Gentzel, Irene N. ^{[1
]}

Park, Chan Ho ^{[1
]}

Bellizzi, Maria ^{[1
]}

Xiao, Guiqing ^{[1
]}

Gadhave, Kiran R. ^{[2
]}

Murphree, Colin ^{[2
]}

Yang, Qin ^{[2
]}

LaMantia, Jonathan ^{[3
]}

Redinbaugh, Margaret G. ^{[1
,3
]}

Balint-Kurti, Peter ^{[2
]}

Sit, Tim L. ^{[2
]}

Wang, Guo-Liang ^{[1
]}

机构：

[1] Ohio State Univ, Dept Plant Pathol, 483B Kottman Hall,2021 Coffey Rd, Columbus, OH 43210 USA

[2] North Carolina State Univ, Dept Entomol & Plant Pathol, Raleigh, NC 27695 USA

[3] ARS, Corn Soybean & Wheat Qual Res Unit, USDA, Wooster, OH 44691 USA

来源：

PLANT METHODS | 2020年 / 16卷 / 01期

关键词：

Maize; Protoplasts; CRISPR; Cas9; Transcription activation; Transcription suppression; AGROBACTERIUM-MEDIATED TRANSFORMATION; MESOPHYLL PROTOPLASTS; MOSAIC-VIRUS; FLORAL DIP; SYSTEM; CRISPR/CAS9; EXPRESSION; PLANT; TRANSCRIPTION; VECTOR;

D O I：

10.1186/s13007-020-00675-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Background TheClusteredRegularlyInterspacedShortPalindromicRepeats (CRISPR)/Cas9 system has become a powerful tool for functional genomics in plants. The RNA-guided nuclease can be used to not only generate precise genomic mutations, but also to manipulate gene expression when present as a deactivated protein (dCas9). Results In this study, we describe a vector toolkit for analyzing dCas9-mediated activation (CRISPRa) or inactivation (CRISPRi) of gene expression in maize protoplasts. An improved maize protoplast isolation and transfection method is presented, as well as a description of dCas9 vectors to enhance or repress maize gene expression. Conclusions We anticipate that this maize protoplast toolkit will streamline the analysis of gRNA candidates and facilitate genetic studies of important trait genes in this transformation-recalcitrant plant.

引用

页数：9

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