ArticleCommunications biology2022
NT-CRISPR, combining natural transformation and CRISPR-Cas9 counterselection for markerless and scarless genome editing in Vibrio natriegens.
Article in Communications biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 56 citations in OpenAlex.
- Metabolic engineering ofSynthetic and systems biotechnology · 2026Article
- Is faster always better? A critical assessment of Vibrio natriegens as microbial host for recombinant protein production.Bioprocess and biosystems engineering · 2026Review
- Mitigating protocatechuic acid toxicity in Vibrio natriegens enables poly-3-hydroxybutyrate production from an aromatic carbon source.Applied microbiology and biotechnology · 2026Article
- Genome editing of phylogenetically distinct bacteria using cross-species retron-mediated recombineering.Nature biotechnology · 2026Article
- CRISPR-Cas based plasmid design for multidrug-resistant Klebsiella pneumoniae isolates.FEMS microbiology letters · 2026Article
- Genetic make-up and regulation of the L-lysine biosynthesis pathway inMicrobial cell (Graz, Austria) · 2026Article
- RECKLEEN is a lambda Red/CRISPR-Cas9 based single plasmid platform for enhanced genome editing in Klebsiella pneumoniae.Communications biology · 2025Article
- Expanding genetic engineering capabilities in Vibrio natriegens with the Vnat Collection.Nucleic acids research · 2025Article
- Article
- A CRISPR-SpCas9M-reporting system for efficient and rapid genome editing in Caulobacter crescentus.Nucleic acids research · 2025Article
- Application of CRISPR-Cas9 in microbial cell factories.Biotechnology letters · 2025Review
- Development of a CRISPR-based cytosine base editor for restriction-modification system inactivation to enhance transformation efficiency in Vibrio Sp. dhg.Journal of biological engineering · 2025Article
- Low-biomass pyruvate production with engineered Vibrio natriegens is accompanied by parapyruvate formation.Microbial cell factories · 2025Article
- Deletion of atypical type II restriction genes in Clostridium cellulovorans using a Cas9-based gene editing system.Applied microbiology and biotechnology · 2025Article
- Vibrio natriegens: Application of a Fast-Growing Halophilic Bacterium.Advances in biochemical engineering/biotechnology · 2025Review
- Molecular mechanisms and applications of natural transformation in bacteria.Frontiers in microbiology · 2025Review
- Protocol for NT-CRISPR: A Method for Efficient Genome Engineering in Vibrio natriegens.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Automation and Miniaturization of Golden Gate DNA Assembly Reactions Using Acoustic Dispensers.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Rationally designed chromosome fusion does not prevent rapid growth of Vibrio natriegens.Communications biology · 2024Article
- CRISPR-Cas9-assisted genome editing inbioRxiv : the preprint server for biology · 2024Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
The fast-growing bacterium Vibrio natriegens has recently gained increasing attention as a novel chassis organism for fundamental research and biotechnology. To fully harness the potential of this bacterium, highly efficient genome editing methods are indispensable to create strains tailored for specific applications. V. natriegens is able to take up free DNA and incorporate it into its genome by homologous recombination. This highly efficient natural transformation is able to mediate uptake of multiple DNA fragments, thereby allowing for multiple simultaneous edits. Here, we describe NT-CRISPR, a combination of natural transformation with CRISPR-Cas9 counterselection. In two temporally distinct steps, we first performed a genome edit by natural transformation and second, induced CRISPR-Cas9 targeting the wild type sequence, and thus leading to death of non-edited cells. Through cell killing with efficiencies of up to 99.999%, integration of antibiotic resistance markers became dispensable, enabling scarless and markerless edits with single-base precision. We used NT-CRISPR for deletions, integrations and single-base modifications with editing efficiencies of up to 100%. Further, we confirmed its applicability for simultaneous deletion of multiple chromosomal regions. Lastly, we showed that the near PAM-less Cas9 variant SpG Cas9 is compatible with NT-CRISPR and thereby broadens the target spectrum.
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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.