Presentation Information
[P03-321]Target-AID base editing facilitates efficient genome modification in lactic acid bacteria
○Hitoshi Mitsunobu1, Keiji Nishida1 (1. Kobe Univ. (Japan))
Keywords:
Base editing,Target-AID,Lactic acid bacteria,Multiplex
Lactic acid bacteria have long played important roles in fermented foods. In recent years, growing interest in their contributions to human health and disease has accelerated their development as probiotics and therapeutic agents. To date, useful strains have mainly been isolated from various natural sources, including the human body, and their genomic and genetic information has been accumulated and made available. While gene function analysis has been advanced using conventional genetic engineering techniques, practical applications in probiotics and food require approaches that do not rely on recombinant DNA.
To address this limitation, we developed a base editing system, Target-AID, optimized for lactic acid bacteria. This system enables the precise introduction of point mutations at multiple genomic loci without the need for donor DNA.
Using this system, we successfully engineered a Lactiplantibacillus plantarum strain with decreased production of imidazole propionate, a metabolite associated with type 2 diabetes. Furthermore, the system enabled multiplex genome editing, allowing simultaneous modification of multiple loci. Transient inactivation of essential genes was also achieved by introducing premature stop codons. Notably, targeting a gene essential for cell division caused severe division defects and induced a filamentous phenotype.
These results indicate that Target-AID enables precise and versatile genome editing in lactic acid bacteria without introducing recombinant DNA, making it suitable for both functional analysis and strain development. Target-AID base editing represents a unique and powerful tool for applications ranging from gene function studies to the generation of beneficial strains in lactic acid bacteria.
To address this limitation, we developed a base editing system, Target-AID, optimized for lactic acid bacteria. This system enables the precise introduction of point mutations at multiple genomic loci without the need for donor DNA.
Using this system, we successfully engineered a Lactiplantibacillus plantarum strain with decreased production of imidazole propionate, a metabolite associated with type 2 diabetes. Furthermore, the system enabled multiplex genome editing, allowing simultaneous modification of multiple loci. Transient inactivation of essential genes was also achieved by introducing premature stop codons. Notably, targeting a gene essential for cell division caused severe division defects and induced a filamentous phenotype.
These results indicate that Target-AID enables precise and versatile genome editing in lactic acid bacteria without introducing recombinant DNA, making it suitable for both functional analysis and strain development. Target-AID base editing represents a unique and powerful tool for applications ranging from gene function studies to the generation of beneficial strains in lactic acid bacteria.
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