ArticleNucleic acids research2014
CRISPR-Cas9-assisted recombineering in Lactobacillus reuteri.
Article in Nucleic acids research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 188 papers.
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Who cites it
188 citing papers in PubMed, 390 citations in OpenAlex.
- Precise probiotic therapy: Advances, bottlenecks, and the road to microbiome-informed nutrition.Gut microbes · 2026Review
- Exploring the emerging role of CRISPR-Cas systems in probiotic development.Engineering microbiology · 2026Review
- Harnessing endogenous CRISPR-Cas9 for inducible genetic engineering ofApplied and environmental microbiology · 2026Article
- An optimized CRISPR/Cas9-based genome editing platform enhances bacteriocin production in Lactiplantibacillus plantarum.Journal of biological engineering · 2026Article
- Development of efficient genetic toolkits and heterologous chassis for bioprospecting of myxobacteria.Nature communications · 2026Article
- Base Editors for Engineering Industrial Microorganisms: Types, Applications, and Future Perspectives.Biotechnology journal · 2026Review
- Ecology and engineering to modify the bile acid output of a defined microbial community.bioRxiv : the preprint server for biology · 2026Article
- Engineering the Gut Microbiome: Emerging Genome-Editing Strategies and Therapeutic Applications.Microorganisms · 2026Review
- Engineered live bacteria for liver diseases and gut-liver axis disorders: from genetic modification to advanced delivery systems.Journal of nanobiotechnology · 2026Review
- Genome editing of phylogenetically distinct bacteria using cross-species retron-mediated recombineering.Nature biotechnology · 2026Article
- Genetically Modified Lactic Acid Bacteria in the EU Food Chain: Applications, Benefits, and Risk Assessment.International journal of molecular sciences · 2026Review
- Application and Research Prospects of CRISPR/Cas Gene Editing Technology in Lactic Acid Bacteria.Microorganisms · 2026Review
- Enhanced editing ofApplied and environmental microbiology · 2026Article
- CRISPR-Cas-Mediated Reprogramming Strategies to Overcome Antimicrobial Resistance.Pharmaceutics · 2026Review
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Search-and-remove genome editing allows selection of cells by DNA sequence.Nature communications · 2025Article
- Harnessing CRISPR-Cas9 for Lactobacillus improvement in silage production: current knowledge and future perspectives.Journal of animal science and biotechnology · 2025Review
- A Cas12a Toolbox for Rapid and Flexible Group B Streptococcus Genomic Editing and CRISPRi.Molecular microbiology · 2025Article
- Chromosomal and Plasmid-Based CRISPRi Platforms for Conditional Gene Silencing inInternational journal of molecular sciences · 2025Article
- Recent advances of engineered probiotics for therapeutic applications.Biodesign research · 2025Review
128 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
Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR-Cas systems, such as the Streptococcus pyogenes CRISPR-Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. Here we have developed and optimized CRISPR-Cas9 function in the lactic acid bacterium Lactobacillus reuteri ATCC PTA 6475. We established proof-of-concept showing that CRISPR-Cas9 selection combined with single-stranded DNA (ssDNA) recombineering is a realistic approach to identify at high efficiencies edited cells in a lactic acid bacterium. We show for three independent targets that subtle changes in the bacterial genome can be recovered at efficiencies ranging from 90 to 100%. By combining CRISPR-Cas9 and recombineering, we successfully applied codon saturation mutagenesis in the L. reuteri chromosome. Also, CRISPR-Cas9 selection is critical to identify low-efficiency events such as oligonucleotide-mediated chromosome deletions. This also means that CRISPR-Cas9 selection will allow identification of recombinant cells in bacteria with low recombineering efficiencies, eliminating the need for ssDNA recombineering optimization procedures. We envision that CRISPR-Cas genome editing has the potential to change the landscape of genome editing in lactic acid bacteria, and other Gram-positive bacteria.
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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.