ArticleNucleic acids research2016
CRISPR-mediated control of the bacterial initiation of replication.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- Elevated temperature undermines the efficacy ofmSphere · 2026Article
- DNA topoisomerase I acts as supercoiling sensor for bacterial transcription elongation.Nature structural & molecular biology · 2026Article
- Harnessing Small RNAs as Synthetic Post-transcriptional Regulators in Bacteria.ACS synthetic biology · 2025Review
- A CRISPR-Cas9 system protecting E. coli against acquisition of antibiotic resistance genes.Scientific reports · 2025Article
- CRISPR-Cas9-driven genome editing inFrontiers in microbiology · 2025Article
- CRISPRi functional genomics in bacteria and its application to medical and industrial research.Microbiology and molecular biology reviews : MMBR · 2024Review
- Article
- Article
- Tips, Tricks, and Potential Pitfalls of CRISPR Genome Editing inFrontiers in bioengineering and biotechnology · 2022Review
- Article
- Review
- A plasmid toolset for CRISPR-mediated genome editing and CRISPRi gene regulation in Escherichia coli.Microbial biotechnology · 2021Article
- Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles.The CRISPR journal · 2021Article
- A catalogue of biochemically diverse CRISPR-Cas9 orthologs.Nature communications · 2020Article
- CRISPR-based gene expression control for synthetic gene circuits.Biochemical Society transactions · 2020Review
- Francisella novicida CRISPR-Cas Systems Can Functionally Complement Each Other in DNA Defense while Providing Target Flexibility.Journal of bacteriology · 2020Article
- CRISPR Tools To Control Gene Expression in Bacteria.Microbiology and molecular biology reviews : MMBR · 2020Review
- Nucleic acid cleavage with a hyperthermophilic Cas9 from an uncultured Ignavibacterium.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Replisome activity slowdown after exposure to ultraviolet light inProceedings of the National Academy of Sciences of the United States of America · 2019Article
- Direct imaging of the circular chromosome in a live bacterium.Nature communications · 2019Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Programmable control of the cell cycle has been shown to be a powerful tool in cell-biology studies. Here, we develop a novel system for controlling the bacterial cell cycle, based on binding of CRISPR/dCas9 to the origin-of-replication locus. Initiation of replication of bacterial chromosomes is accurately regulated by the DnaA protein, which promotes the unwinding of DNA at oriC We demonstrate that the binding of CRISPR/dCas9 to any position within origin or replication blocks the initiation of replication. Serial-dilution plating, single-cell fluorescence microscopy, and flow-cytometry experiments show that ongoing rounds of chromosome replication are finished upon CRISPR/dCas9 binding, but no new rounds are initiated. Upon arrest, cells stay metabolically active and accumulate cell mass. We find that elevating the temperature from 37 to 42°C releases the CRISR/dCas9 replication inhibition, and we use this feature to recover cells from the arrest. Our simple and robust method of controlling the bacterial cell cycle is a useful asset for synthetic biology and DNA-replication studies in particular. The inactivation of CRISPR/dCas9 binding at elevated temperatures may furthermore be of wide interest for CRISPR/Cas9 applications in genomic engineering.
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Registered trials
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