期刊
PLANT METHODS
卷 16, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s13007-020-00675-5
关键词
Maize; Protoplasts; CRISPR; Cas9; Transcription activation; Transcription suppression
资金
- DARPA's Insect Allies Program
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.
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