ArticleMicrobial cell factories2018
Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system.
Article in Microbial cell factories, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed.
- High-level prodigiosin production inSynthetic and systems biotechnology · 2026Article
- A portable Cas6f-based system for multiplex translational repression in bacteria.Nature communications · 2026Article
- Resilience of recombinant antibiotic resistance gene-containing plasmids against common cell culture disposal methods.Cell reports methods · 2026Article
- Base Editors for Engineering Industrial Microorganisms: Types, Applications, and Future Perspectives.Biotechnology journal · 2026Review
- Exploiting the Endogenous Type II-A CRISPR-Cas System for Functional Engineering of Probiotic Lacticaseibacillus rhamnosus GG.Microbial biotechnology · 2026Article
- Transcriptome-guided discovery of novel plant-associated genes in a rhizosphere Pseudomonas.Microbiome · 2025Article
- Programming Nutrient Detection with Modular Regulators for Dynamic Control of Microbial Biosynthesis.ACS synthetic biology · 2025Article
- A single-plasmid-based, easily curable CRISPR/Cas9 system for rapid, iterative genome editing in Pseudomonas putida KT2440.Microbial cell factories · 2024Article
- Modification of Glucose Metabolic Pathway to Enhance Polyhydroxyalkanoate Synthesis inCurrent issues in molecular biology · 2024Article
- Establishing a straightforward I-SceI-mediated recombination one-plasmid system for efficient genome editing in P. putida KT2440.Microbial biotechnology · 2024Article
- Genomic editing inApplied and environmental microbiology · 2024Article
- Cyclic di-GMP inhibits nitrate assimilation by impairing the antitermination function of NasT in Pseudomonas putida.Nucleic acids research · 2024Article
- Computational design of CRISPR guide RNAs to enable strain-specific control of microbial consortia.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Dynamic and single cell characterization of a CRISPR-interference toolset inMetabolic engineering communications · 2022Article
- Highly efficient genome editing in Xanthomonas oryzae pv. oryzae through repurposing the endogenous type I-C CRISPR-Cas system.Molecular plant pathology · 2022Article
- CRISPR-Based Approaches for Gene Regulation in Non-Model Bacteria.Frontiers in genome editing · 2022Review
- Streamlined CRISPR genome engineering in wild-type bacteria using SIBR-Cas.Nucleic acids research · 2021Article
- Development of dual-inducible duet-expression vectors for tunable gene expression control and CRISPR interference-based gene repression in Pseudomonas putida KT2440.Microbial biotechnology · 2021Article
- Genome Editing in Bacteria: CRISPR-Cas and Beyond.Microorganisms · 2021Review
- CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering.Nature biotechnology · 2021Article
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7 authors.
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Abstract
backgroundThe soil bacterium Pseudomonas putida KT2440 is a "generally recognized as safe"-certified strain with robust property and versatile metabolism. Thus, it is an ideal candidate for synthetic biology, biodegradation, and other biotechnology applications. The known genome editing approaches of Pseudomonas are suboptimal; thus, it is necessary to develop a high efficiency genome editing tool.
resultsIn this study, we established a fast and convenient CRISPR-Cas9 method in P. putida KT2440. Gene deletion, gene insertion and gene replacement could be achieved within 5 days, and the mutation efficiency reached > 70%. Single nucleotide replacement could be realized, overcoming the limitations of protospacer adjacent motif sequences. We also applied nuclease-deficient Cas9 binding at three locations upstream of enhanced green fluorescent protein (eGFP) for transcriptional inhibition, and the expression intensity of eGFP reduced to 28.5, 29.4, and 72.1% of the control level, respectively. Furthermore, based on this CRISPR-Cas9 system, we also constructed a CRISPR-Cpf1 system, which we validated for genome editing in P. putida KT2440.
conclusionsIn this research, we established CRISPR based genome editing and regulation control systems in P. putida KT2440. These fast and efficient approaches will greatly facilitate the application of P. putida KT2440.
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