ArticleMicrobial biotechnology2020
Accelerated genome engineering of Pseudomonas putida by I-SceI-mediated recombination and CRISPR-Cas9 counterselection.
Article in Microbial biotechnology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 75 papers.
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Who cites it
75 citing papers in PubMed, 153 citations in OpenAlex.
- Growth-coupled microbial biosynthesis of the animal pigment xanthommatin.Nature biotechnology · 2026Article
- A portable Cas6f-based system for multiplex translational repression in bacteria.Nature communications · 2026Article
- Cascading recombinase memory switch for programmable and stable gene expression in Pseudomonas putida.Nucleic acids research · 2026Article
- Ethyl methanesulfonate mutagenesis: Advancing bacterial genetics for sustainable agriculture.Folia microbiologica · 2026Review
- Adaptive evolution ofJournal of bacteriology · 2026Article
- On the Choice of the Right Plasmid Vector(s) in the Times of Synthetic Biology.Microbial biotechnology · 2025Review
- Combinatorial engineering pinpoints shikimate pathway bottlenecks in para-aminobenzoic acid production in Pseudomonas putida.Journal of biological engineering · 2025Article
- Synthetic CmBio · 2025Article
- The TonB-Dependent Transport System Facilitates the Uptake of Inorganic Metal Mediators in Pseudomonas putida KT2440 in a Bioelectrochemical System.Microbial biotechnology · 2025Article
- Evaluation of Engineering Potential in Undomesticated Microbes With VECTOR.Microbial biotechnology · 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
- Release of extracellular DNA byiScience · 2025Article
- Engineering a phi15-based expression system for stringent gene expression in Pseudomonas putida.Communications biology · 2025Article
- Leveraging EngineeredACS synthetic biology · 2025Article
- Presence of a biofilm beneficiary alters the evolutionary trajectory of a biofilm former.The ISME journal · 2025Article
- Construction of a syntrophicSynthetic biology (Oxford, England) · 2025Article
- Visualization of Type IV-A1 CRISPR-mediated repression of gene expression and plasmid replication.Nucleic acids research · 2024Article
- Enhanced biosynthesis of poly(3-hydroxybutyrate) in engineered strains of Pseudomonas putida via increased malonyl-CoA availability.Microbial biotechnology · 2024Article
- A versatile microbial platform as a tunable whole-cell chemical sensor.Nature communications · 2024Article
15 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Pseudomonas species have become reliable platforms for bioproduction due to their capability to tolerate harsh conditions imposed by large-scale bioprocesses and their remarkable resistance to diverse physicochemical stresses. The last few years have brought forth a variety of synthetic biology tools for the genetic manipulation of pseudomonads, but most of them are either applicable only to obtain certain types of mutations, lack efficiency, or are not easily accessible to be used in different Pseudomonas species (e.g. natural isolates). In this work, we describe a versatile, robust and user-friendly procedure that facilitates virtually any kind of genomic manipulation in Pseudomonas species in 3-5 days. The protocol presented here is based on DNA recombination forced by double-stranded DNA cuts (through the activity of the I-SceI homing meganuclease from yeast) followed by highly efficient counterselection of mutants (aided by a synthetic CRISPR-Cas9 device). The individual parts of the genome engineering toolbox, tailored for knocking genes in and out, have been standardized to enable portability and easy exchange of functional gene modules as needed. The applicability of the procedure is illustrated both by eliminating selected genomic regions in the platform strain P. putida KT2440 (including difficult-to-delete genes) and by integrating different reporter genes (comprising novel variants of fluorescent proteins) into a defined landing site in the target chromosome.
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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.