CRISPR-assisted multi-dimensional regulation for fine-tuning gene expression in Bacillus subtilis.

ABSTRACT: Fine-tuning of gene expression is crucial for protein expression and pathway construction, but it still faces formidable challenges due to the hierarchical gene regulation at multiple levels in a context-dependent manner. In this study, we defined the optimal targeting windows for CRISPRa and CRISPRi of the dCas9-α/ω system, and demonstrated that this system could act as a single master regulator to simultaneously activate and repress the expression of different genes by designing position-specific gRNAs. The application scope of dCas9-ω was further expanded by a newly developed CRISPR-assisted Oligonucleotide Annealing based Promoter Shuffling (OAPS) strategy, which could generate a high proportion of functional promoter mutants and facilitate the construction of effective promoter librar

SUBMITTER: Lu Z 

PROVIDER: S-EPMC6468239 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature