ArticleNucleic acids research2016
Consequences of Cas9 cleavage in the chromosome of Escherichia coli.
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 149 papers.
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Who cites it
149 citing papers in PubMed, 287 citations in OpenAlex.
- Technical limitations of CRISPR-Cas9 genome editing in bacteria: challenges and future directions.Archives of microbiology · 2026Review
- Phage and CRISPR based precision antimicrobials: a dual strategy against multidrug-resistant bacteria.Molecular biology reports · 2026Review
- Probing the limits of genetic recoding using multi-omics-guided evolution.Nature communications · 2026Article
- Inducible CRISPRi enables efficient and high-fidelity genome editing in Streptomyces.Nucleic acids research · 2026Article
- RECC: A Red/ET-CRISPR/Cas9-based system enabling genome mining of marineSynthetic and systems biotechnology · 2026Article
- Multiplexed CRISPR base editing enables pulse-activated irreversible biocontainment of engineered bacteria.Nucleic acids research · 2026Article
- Forecasting Multitrait Resistance Evolution under Antibiotic Stress.Molecular biology and evolution · 2026Article
- A conjugal gene drive-like system efficiently suppresses antibiotic resistance in a bacterial population.npj antimicrobials and resistance · 2026Article
- Bacteria as living biosensors for DNA.Nature reviews bioengineering · 2026Article
- Multilayered regulation by RNA thermometers enables precise control of Cas9 expression in E. coli.Nucleic acids research · 2026Article
- Machine learning reveals sequence and methylation determinants of SaCas9-PAM interactions in bacteria.Nucleic acids research · 2026Article
- Specific killing and resensitization of pathogenic Escherichia coli strains carrying blaCTX-M-15 β-lactamase using targeted-antibacterial-plasmids (TAPs).Nucleic acids research · 2026Article
- Avian PathogenicVeterinary sciences · 2025Review
- Tailoring Cas12a functionality with a user-friendly and versatile crRNA variant toolbox.Nature communications · 2025Article
- Genetic Entanglement Enables Ultrastable Biocontainment in the Mammalian Gut.ACS synthetic biology · 2025Article
- Controlling CRISPR-Cas9 genome editing in human cells using a molecular glue degrader.Molecular therapy. Nucleic acids · 2025Article
- EASY-edit: a toolbox for high-throughput single-step custom genetic editing in bacteria.Nucleic acids research · 2025Article
- Phage susceptibility to a minimal, modular synthetic CRISPR-Cas system inPhilosophical transactions of the Royal Society of London. Series B, Biological sciences · 2025Article
- Conjugative delivery of toxin genesJournal of bacteriology · 2025Article
- Applications of multiplexed CRISPR-Cas for genome engineering.Experimental & molecular medicine · 2025Review
89 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The RNA-guided Cas9 nuclease from CRISPR-Cas systems has emerged as a powerful biotechnological tool. The specificity of Cas9 can be reprogrammed to cleave desired sequences in a cell's chromosome simply by changing the sequence of a small guide RNA. Unlike in most eukaryotes, Cas9 cleavage in the chromosome of bacteria has been reported to kill the cell. However, the mechanism of cell death remains to be investigated. Bacteria mainly rely on homologous recombination (HR) with sister chromosomes to repair double strand breaks. Here, we show that the simultaneous cleavage of all copies of the Escherichia coli chromosome at the same position cannot be repaired, leading to cell death. However, inefficient cleavage can be tolerated through continuous repair by the HR pathway. In order to kill cells reliably, HR can be blocked using the Mu phage Gam protein. Finally, the introduction of the non-homologous end joining (NHEJ) pathway from Mycobacterium tuberculosis was not able to rescue the cells from Cas9-mediated killing, but did introduce small deletions at a low frequency. This work provides a better understanding of the consequences of Cas9 cleavage in bacterial chromosomes which will be instrumental in the development of future CRISPR tools.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.