ArticleNucleic acids research2020
Design of a programmable biosensor-CRISPRi genetic circuits for dynamic and autonomous dual-control of metabolic flux in Bacillus subtilis.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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57 citing papers in PubMed, 173 citations in OpenAlex.
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- Advances in natural product discovery: strategies, technologies, and insights.Natural products and bioprospecting · 2026Review
- Optimization Strategy of Expression Vectors and Regulatory Elements for Enhanced Protein Production inInternational journal of molecular sciences · 2025Review
- Characterizing and Engineering a Succinate-Responsive Biosensor System inACS synthetic biology · 2025Article
- Robust production of N-acetyl-glucosamine in engineeredSynthetic and systems biotechnology · 2025Article
- A quorum sensing-controlled type I CRISPRi toolkit for dynamically regulating metabolic flux.Nucleic acids research · 2025Article
- Epigenome Engineering Using dCas Systems for Biomedical Applications and Biotechnology: Current Achievements, Opportunities and Challenges.International journal of molecular sciences · 2025Review
- Combining biosensor and metabolic network optimization strategies for enhanced L-threonine production in Escherichia coli.Biotechnology for biofuels and bioproducts · 2025Article
- A cross-species inducible system for enhanced protein expression and multiplexed metabolic pathway fine-tuning in bacteria.Nucleic acids research · 2025Article
- Optogenetic control of Corynebacterium glutamicum gene expression.Nucleic acids research · 2024Article
- Synthetic gene circuit evolution: Insights and opportunities at the mid-scale.Cell chemical biology · 2024Review
- Dynamic and Static Regulation of Nicotinamide Adenine Dinucleotide Phosphate: Strategies, Challenges, and Future Directions in Metabolic Engineering.Molecules (Basel, Switzerland) · 2024Review
- Investigating and Engineering an 1,2-Propanediol-Responsive Transcription Factor-Based Biosensor.ACS synthetic biology · 2024Article
- Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria.Nature communications · 2024Article
- Regulatory RNAs inSynthetic and systems biotechnology · 2024Review
- Reshaping Phosphatase Substrate Preference for Controlled Biosynthesis Using a "Design-Build-Test-Learn" Framework.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor.Nucleic acids research · 2024Article
- CRISPR in Nucleic Acids Research: the sequel.Nucleic acids research · 2024Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Dynamic regulation is an effective strategy for fine-tuning metabolic pathways in order to maximize target product synthesis. However, achieving dynamic and autonomous up- and down-regulation of the metabolic modules of interest simultaneously, still remains a great challenge. In this work, we created an autonomous dual-control (ADC) system, by combining CRISPRi-based NOT gates with novel biosensors of a key metabolite in the pathway of interest. By sensing the levels of the intermediate glucosamine-6-phosphate (GlcN6P) and self-adjusting the expression levels of the target genes accordingly with the GlcN6P biosensor and ADC system enabled feedback circuits, the metabolic flux towards the production of the high value nutraceutical N-acetylglucosamine (GlcNAc) could be balanced and optimized in Bacillus subtilis. As a result, the GlcNAc titer in a 15-l fed-batch bioreactor increased from 59.9 g/l to 97.1 g/l with acetoin production and 81.7 g/l to 131.6 g/l without acetoin production, indicating the robustness and stability of the synthetic circuits in a large bioreactor system. Remarkably, this self-regulatory methodology does not require any external level of control such as the use of inducer molecules or switching fermentation/environmental conditions. Moreover, the proposed programmable genetic circuits may be expanded to engineer other microbial cells and metabolic pathways.
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