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A CRISPR/dCas9 toolkit for functional analysis of maize genes.

ABSTRACT:

Background

The Clustered Regularly Interspaced Short Palindromic Repeats (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.

SUBMITTER: Gentzel IN

PROVIDER: S-EPMC7532566 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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A CRISPR/dCas9 toolkit for functional analysis of maize genes.

Gentzel Irene N IN Park Chan Ho CH Bellizzi Maria M Xiao Guiqing G Gadhave Kiran R KR Murphree Colin C Yang Qin Q LaMantia Jonathan J Redinbaugh Margaret G MG Balint-Kurti Peter P Sit Tim L TL Wang Guo-Liang GL

Plant methods 20201002

<h4>Background</h4>The Clustered Regularly Interspaced Short Palindromic Repeats (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).<h4>Results</h4>In this study, we describe a vector toolkit for analyzing dCas9-mediated activation (CRISPRa) or inactivation (CRISPRi) of gene expression in maize pro ...[more]

PMID: 33024447

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