ArticleNucleic acids research2019
CRISPR-assisted multi-dimensional regulation for fine-tuning gene expression in Bacillus subtilis.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed.
- Multi-parameter optimization of recombinant protein secretion in Bacillus subtilis: a review.Microbial cell factories · 2026Review
- Development of Bacillus subtilis cell factories for nutraceuticals production.World journal of microbiology & biotechnology · 2026Review
- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- Regulation of Riboflavin Biosynthesis in Microorganisms and Construction of the Advanced Overproducers of This Vitamin.International journal of molecular sciences · 2025Review
- Identification of ice-binding proteins from Raphanus sativus and application in frozen dough.NPJ science of food · 2025Article
- CRISPR/Cas13X-assisted programmable and multiplexed translation regulation for controlled biosynthesis.Nucleic acids research · 2025Article
- CRISPR/Cas9-Engineering for Increased Amylolytic Potential of Microbes for Sustainable Wastewater Treatment: A Review.Current microbiology · 2024Review
- Design and Characterization of a Transcriptional Repression Toolkit for Plants.ACS synthetic biology · 2024Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- Genome-Wide CRISPRi Screening of Key Genes for Recombinant Protein Expression in Bacillus Subtilis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Biosensor-assisted CRISPRi high-throughput screening to identify genetic targets inSynthetic and systems biotechnology · 2024Article
- Toolbox of Characterized Genetic Parts forACS synthetic biology · 2024Article
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- Engineered bacterial orthogonal DNA replication system for continuous evolution.Nature chemical biology · 2023Article
- Novel epigenetic molecular therapies for imprinting disorders.Molecular psychiatry · 2023Review
- CRISPR-dCas12a-mediated genetic circuit cascades for multiplexed pathway optimization.Nature chemical biology · 2023Article
- Article
- Barriers to simultaneous multilocus integration in Bacillus subtilis tumble down: development of a straightforward screening method for the colorimetric detection of one-step multiple gene insertion using the CRISPR-Cas9 system.Microbial cell factories · 2023Article
- Recent Advances in CRISPR-Cas Technologies for Synthetic Biology.Journal of microbiology (Seoul, Korea) · 2023Review
- Construction and application of an efficient dual-base editing platform forChemical science · 2022Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
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 libraries in microorganisms with low transformation efficiency. Combing OAPS and dCas9-ω, the influences of promoter-based transcription, molecular chaperone-assisted protein folding and protease-mediated degradation on the expression of amylase BLA in Bacillus subtilis were systematically evaluated, and a 260-fold enhancement of BLA production was obtained. The success of the OAPS strategy and dCas9-ω for BLA production in this study thus demonstrated that it could serve as a powerful tool kit to regulate the expression of multiple genes multi-directionally and multi-dimensionally in bacteria.
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