ArticleNucleic acids research2023
CRISPRi-mediated tunable control of gene expression level with engineered single-guide RNA in Escherichia coli.
Article in Nucleic acids research, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Toward engineered microbial chassis for growth-production balancing.Synthetic and systems biotechnology · 2027Review
- Programmable in vivo mRNA circularization for enhanced gene expression in bacteria.Nucleic acids research · 2026Article
- Biosynthesis and Microbial Production of Carminic Acid: From Pathway Elucidation to Synthetic Biology.Microorganisms · 2026Review
- Engineering Extracellular Vesicles for Anti-Aging Therapy: Mechanisms, Applications, and Perspectives.Aging cell · 2026Review
- Multiplexed CRISPR base editing enables pulse-activated irreversible biocontainment of engineered bacteria.Nucleic acids research · 2026Article
- Coordinating Calvin Cycle and Glycolysis inACS synthetic biology · 2026Article
- Systematic CRISPRi screening reveals genetic modulators ofbioRxiv : the preprint server for biology · 2026Article
- Synthetic Biology Strategies for Activating Cryptic BGCs inACS omega · 2026Review
- Design of Conditional Guide RNAs for the Logical Regulation of Gene Expression.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Tailoring Cas12a functionality with a user-friendly and versatile crRNA variant toolbox.Nature communications · 2025Article
- Considering Metabolic Context in Enzyme Evolution and Design.Biochemistry · 2025Review
- Design of function-regulating RNA via deep learning and AlphaFold 3.Briefings in bioinformatics · 2025Article
- Generation and validation of a versatile inducible multiplex CRISPRi system to examine bacterial regulation in the Euprymna-Vibrio fischeri symbiosis.Archives of microbiology · 2025Article
- Dynamic control of the plasmid copy number maintained without antibiotics in Escherichia coli.Journal of biological engineering · 2024Article
- Guide RNA structure design enables combinatorial CRISPRa programs for biosynthetic profiling.Nature communications · 2024Article
- Pigments from pathogenic bacteria: a comprehensive update on recent advances.World journal of microbiology & biotechnology · 2024Review
- CRISPRi functional genomics in bacteria and its application to medical and industrial research.Microbiology and molecular biology reviews : MMBR · 2024Review
- Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria.Nature communications · 2024Article
- CRISPRi: a way to integrate iPSC-derived neuronal models.Biochemical Society transactions · 2024Review
- Advances and applications of CRISPR/Cas-mediated interference inEngineering microbiology · 2024Review
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Precise control of gene expression is essential for flux redistribution in metabolic pathways. Although the CRISPR interference (CRISPRi) system can effectively repress gene expression at the transcriptional level, it has still been difficult to precisely control the level without loss of specificity or an increase in cell toxicity. In this study, we developed a tunable CRISPRi system that performs transcriptional regulation at various levels. We constructed a single-guide RNA (sgRNA) library targeting repeat, tetraloop, and anti-repeat regions to modulate the binding affinity against dCas9. Each screened sgRNA could regulate the gene expression at a certain level between fully-repressing and non-repressing states (>45-fold). These sgRNAs also enabled modular regulation with various target DNA sequences. We applied this system to redistribute the metabolic flux to produce violacein derivatives in a predictable ratio and optimize lycopene production. This system would help accelerate the flux optimization processes in metabolic engineering and synthetic biology.
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