ArticleNature communications2022
Modular (de)construction of complex bacterial phenotypes by CRISPR/nCas9-assisted, multiplex cytidine base-editing.
Article in Nature communications, 2022. 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.
- A portable Cas6f-based system for multiplex translational repression in bacteria.Nature communications · 2026Article
- Mind the Porins: Differential Effects of Porin Knockouts and Overexpression on Glucose and Xylose Uptake and Utilization in Pseudomonas putida.Microbial biotechnology · 2026Article
- Cascading recombinase memory switch for programmable and stable gene expression in Pseudomonas putida.Nucleic acids research · 2026Article
- Genome editing of phylogenetically distinct bacteria using cross-species retron-mediated recombineering.Nature biotechnology · 2026Article
- Zinc starvation uncovers bacterial host-specific proteases that shape NDM adaptability inmBio · 2026Article
- Characterization and engineering of a type IV-A3 CRISPR-Cas system for genome editing in Escherichia coli.Nucleic acids research · 2025Article
- Synthetic CmBio · 2025Article
- Transcriptomics ofMicrobiology resource announcements · 2025Article
- CRISPR-driven enhanced hydrocarbon emulsification in an environmental Pseudomonas aeruginosa strain.Microbial cell factories · 2025Article
- Genetic Dissection of Cyclic di-GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption.Microbial biotechnology · 2025Article
- Computation-aided designs enable developing auxotrophic metabolic sensors for wide-range glyoxylate and glycolate detection.Nature communications · 2025Article
- Upcycling of polyamides through chemical hydrolysis and engineered Pseudomonas putida.Nature microbiology · 2025Article
- Applications of bacteriophages in precision engineering of the human gut microbiome.Engineering microbiology · 2025Review
- Precision engineering of the probioticApplied and environmental microbiology · 2025Article
- Engineering a phi15-based expression system for stringent gene expression in Pseudomonas putida.Communications biology · 2025Article
- A universal and wide-range cytosine base editor via domain-inlaid and fidelity-optimized CRISPR-FrCas9.Nature communications · 2025Article
- Leveraging EngineeredACS synthetic biology · 2025Article
- Hypermutability bypasses genetic constraints in SCV phenotypic switching in Pseudomonas aeruginosa biofilms.NPJ biofilms and microbiomes · 2025Article
- Selection of Fusion-Site Overhang Sets for High-Fidelity and High-Complexity Golden Gate Assembly.Methods in molecular biology (Clifton, N.J.) · 2025Article
- One-for-all gene inactivation via PAM-independent base editing in bacteria.The Journal of biological chemistry · 2025Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas technologies constitute a powerful tool for genome engineering, yet their use in non-traditional bacteria depends on host factors or exogenous recombinases, which limits both efficiency and throughput. Here we mitigate these practical constraints by developing a widely-applicable genome engineering toolset for Gram-negative bacteria. The challenge is addressed by tailoring a CRISPR base editor that enables single-nucleotide resolution manipulations (C·G → T·A) with >90% efficiency. Furthermore, incorporating Cas6-mediated processing of guide RNAs in a streamlined protocol for plasmid assembly supports multiplex base editing with >85% efficiency. The toolset is adopted to construct and deconstruct complex phenotypes in the soil bacterium Pseudomonas putida. Single-step engineering of an aromatic-compound production phenotype and multi-step deconstruction of the intricate redox metabolism illustrate the versatility of multiplex base editing afforded by our toolbox. Hence, this approach overcomes typical limitations of previous technologies and empowers engineering programs in Gram-negative bacteria that were out of reach thus far.
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