2020 journal article

A CRISPR/dCas9 toolkit for functional analysis of maize genes

PLANT METHODS, 16(1).

By: I. Gentzel*, C. Park*, M. Bellizzi*, G. Xiao*, K. Gadhave n, C. Murphree n, Q. Yang n, J. LaMantia* ...

co-author countries: United States of America 🇺🇸
author keywords: Maize; Protoplasts; CRISPR; Cas9; Transcription activation; Transcription suppression
Source: Web Of Science
Added: October 26, 2020

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).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.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.