ArticleBioengineered bugs
Efficacy of a Lactococcus lactis ΔpyrG vaccine delivery platform expressing chromosomally integrated hly from Listeria monocytogenes.
Article in Bioengineered bugs. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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11 citing papers in PubMed, 30 citations in OpenAlex.
- Quadruplet codon decoding-based versatile genetic biocontainment system.Nucleic acids research · 2025Article
- Biocontainment strategies forFrontiers in bioengineering and biotechnology · 2023Article
- Microbial Biocontainment Systems for Clinical, Agricultural, and Industrial Applications.Frontiers in bioengineering and biotechnology · 2022Review
- Novel Strategies for Efficient Production and Delivery of Live Biotherapeutics and Biotechnological Uses ofFrontiers in bioengineering and biotechnology · 2020Review
- Fail-safe genetic codes designed to intrinsically contain engineered organisms.Nucleic acids research · 2019Article
- Development of bacteria as diagnostics and therapeutics by genetic engineering.Journal of microbiology (Seoul, Korea) · 2019Review
- Next-generation biocontainment systems for engineered organisms.Nature chemical biology · 2018Article
- Listeria monocytogenes mutants defective in gallbladder replication represent safety-enhanced vaccine delivery platforms.Human vaccines & immunotherapeutics · 2016Article
- Designer probiotics: Development and applications in gastrointestinal health.World journal of gastrointestinal pathophysiology · 2015Review
- Two-tiered biological containment strategy for Lactococcus lactis-based vaccine or immunotherapy vectors.Human vaccines & immunotherapeutics · 2014Article
- Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Listeria monocytogenes is a significant food-borne pathogen and the causative agent of listeriosis, a disease which manifests as meningitis in immunocompromised adults or infection of the fetus and miscarriage in pregnant women. We have previously used Lactococcus lactis, a GRAS (Generally Regarded As Safe) organism, as a vaccine vector against listeriosis by engineering plasmid-mediated expression of the immunodominant antigen from L. monocytogenes, listeriolysin O (LLO). However, the environmental release of an engineered vaccine vector carrying a replicating plasmid during clinical usage may raise safety concerns. Here we describe the integration of the LLO gene (hly) into the L. lactis chromosome through homologous double crossover to allow stable expression, in order to avoid the use of antibiotic selection markers and to eliminate the requirement for a plasmid-based system. The approach was designed to simultaneously eliminate the pyrG gene encoding the CTP synthase which is responsible for converting UTP to CTP in a unique step in the de novo pyrimidine synthesis in L. lactis. This gene was targeted in order to restrict bacterial replication outside of the host (biological containment). The resulting cytidine auxotroph was able to secrete LLO constitutively and could elicit LLO(91-99)-specific CD8(+) T lymphocytes in the murine infection model. Moreover, protection against lethal challenge with L. monocytogenes was accomplished after intraperitoneal (IP) vaccination with the constructed strain. The implications for the use of cytidine auxotropy in biological containment are discussed.
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