ArticleScientific reports2018
Single plasmid systems for inducible dual protein expression and for CRISPR-Cas9/CRISPRi gene regulation in lactic acid bacterium Lactococcus lactis.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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55 citing papers in PubMed, 105 citations in OpenAlex.
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- An optimized CRISPR/Cas9-based genome editing platform enhances bacteriocin production in Lactiplantibacillus plantarum.Journal of biological engineering · 2026Article
- Prospects and challenges in using engineered lactic acid bacteria in aquaculture applications.Engineering microbiology · 2026Review
- Multienzyme Platform for the Synthesis of UDP Sugars and Human Milk Oligosaccharides.Chembiochem : a European journal of chemical biology · 2026Article
- Application and Research Prospects of CRISPR/Cas Gene Editing Technology in Lactic Acid Bacteria.Microorganisms · 2026Review
- Enhanced editing ofApplied and environmental microbiology · 2026Article
- EngineeringFrontiers in bioengineering and biotechnology · 2026Article
- Probiotic-Based Materials as Living Therapeutics.Advanced materials (Deerfield Beach, Fla.) · 2026Review
- Being a better version of yourself: genetically engineered probiotic bacteria as host defense enhancers in the control of intestinal pathogens.Gut microbes · 2025Review
- Chromosomal and Plasmid-Based CRISPRi Platforms for Conditional Gene Silencing inInternational journal of molecular sciences · 2025Article
- Unlocking Casein Bioactivity: Lactic Acid Bacteria and Molecular Strategies for Peptide Release.International journal of molecular sciences · 2025Review
- Harnessing the Microbiome: CRISPR-Based Gene Editing and Antimicrobial Peptides in Combating Antibiotic Resistance and Cancer.Probiotics and antimicrobial proteins · 2025Review
- Cryo-EM analyses unveil details of mechanism and targocil-II mediated inhibition of S. aureus WTA transporter TarGH.Nature communications · 2025Article
- Gut Microbiota Modulation in IBD: From the Old Paradigm to Revolutionary Tools.International journal of molecular sciences · 2025Review
- A Plasmid-Encoded Surface Polysaccharide Partly BlocksInternational journal of molecular sciences · 2025Article
- Incorporation of recombinant proteins into extracellular vesicles by Lactococcus cremoris.Scientific reports · 2025Article
- Application of the SpCas9 inhibitor BRD0539 for CRISPR/Cas9-based genetic tools inBioscience of microbiota, food and health · 2025Article
- Next-generation probiotics and engineered BEVs for precision therapeutics in osteoporosis.Frontiers in nutrition · 2025Review
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Lactococcus lactis is a food-grade lactic acid bacterium that is used in the dairy industry as a cell factory and as a host for recombinant protein expression. The nisin-controlled inducible expression (NICE) system is frequently applied in L. lactis; however new tools for its genetic modification are highly desirable. In this work NICE was adapted for dual protein expression. Plasmid pNZDual, that contains two nisin promoters and multiple cloning sites (MCSs), and pNZPolycist, that contains a single nisin promoter and two MCSs separated by the ribosome binding site, were constructed. Genes for the infrared fluorescent protein and for the human IgG-binding DARPin were cloned in all possible combinations to assess the protein yield. The dual promoter plasmid pNZDual enabled balanced expression of the two model proteins. It was exploited for the development of a single-plasmid inducible CRISPR-Cas9 system (pNZCRISPR) by using a nisin promoter, first to drive Cas9 expression and, secondly, to drive single guide RNA transcription. sgRNAs against htrA and ermR directed Cas9 against genomic or plasmid DNA and caused changes in bacterial growth and survival. Replacing Cas9 by dCas9 enabled CRISPR interference-mediated silencing of the upp gene. The present study introduces a new series of plasmids for advanced genetic modification of lactic acid bacterium L. lactis.
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