Hyper-Blebbing Bacteria via rpsA Gene Modification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing native outer membrane vesicles (nOMV) from Gram-negative bacteria are limited by the variability of hyper-blebbing mutations across different bacterial species, making it challenging to induce hyper-blebbing in all Gram-negative bacteria, which is essential for large-scale vaccine production.
Innovation Solution
Modifying the rpsA gene, rpsA operon, and/or 30S ribosomal protein S1 to increase nOMV release, as these modifications are broadly conserved across bacteria genera and species, allowing for increased nOMV production without disrupting membrane structural components.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If deletion of genes encoding key membrane structural components (such as gna33 or tolR) is performed to achieve hyper-blebbing phenotype, then larger quantities of outer membrane vesicles are produced, but the method is not universally applicable across different Gram-negative bacterial species due to genetic diversity and species-specific biochemical mechanisms
Solution Approach 1:
The patent applies universality by targeting the rpsA gene, which is highly conserved across all Gram-negative bacterial species. By modifying this universally present gene rather than species-specific membrane structural genes, the hyper-blebbing phenotype can be induced across diverse Gram-negative bacteria including E. coli, Salmonella, Pseudomonas, and others, making the method universally applicable while maintaining high nOMV production quantities
Solution Approach 2:
The patent employs parameter changes by modifying the rpsA gene sequence or expression levels to alter ribosomal protein S1 function. This genetic parameter modification triggers a phenotypic change (hyper-blebbing) that increases nOMV production, and since rpsA is conserved across species, the same parameter modification strategy can be applied universally to different Gram-negative bacteria
2Adaptability or versatility
If random mutation and testing is performed to screen for hyper-blebbing mutations in new bacterial species, then potential hyper-blebbing mutations can be identified, but the process is time-consuming and requires functional screening for each species
Solution Approach 1:
The patent applies preliminary action by identifying and validating the rpsA gene as a universal target for hyper-blebbing induction before applying the method to new bacterial species. Since rpsA conservation has been established, the preliminary work of screening and validation has already been done, allowing direct application to new species without time-consuming random mutation and functional screening processes
Data Source
AI summary
The present invention relates to the field of hyper-blebbing Gram-negative bacterial cells which are genetically modified by modifying the rpsA gene, the rpsA operon and/or 30S ribosomal protein S1 and to native outer membrane vesicles (nOMVs) obtained or obtainable from said genetically modified bacterial cells.