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 facili

SUBMITTER: Gentzel IN 

PROVIDER: S-EPMC7532566 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature