A CRISPR/dCas9 toolkit for functional analysis of maize genes.
- Additional Document Info
- View All
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.
author list (cited authors)
Gentzel, I. N., Park, C. H., Bellizzi, M., Xiao, G., Gadhave, K. R., Murphree, C., ... Wang, G.
complete list of authors
Gentzel, Irene N||Park, Chan Ho||Bellizzi, Maria||Xiao, Guiqing||Gadhave, Kiran R||Murphree, Colin||Yang, Qin||LaMantia, Jonathan||Redinbaugh, Margaret G||Balint-Kurti, Peter||Sit, Tim L||Wang, Guo-Liang