AI Article Synopsis
- The CRISPR/Cas9 gene editing system has been effectively used in various organisms, but applications in lactic acid bacteria are still in early stages.
- A new recombinant system combining CRISPR/Cas9 with Red/ET technology was developed using a plasmid from Lactococcus lactis, showing a knockout efficiency of 91%.
- This efficient system avoids the use of antibiotics or exogenous genes and offers high-throughput screening potential for gene editing in lactic acid bacteria.
Article Abstract
Clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system has been widely used in gene editing of various organisms. However, food-grade gene editing systems in lactic acid bacteria are still preliminary. Red/ET-dependent homologous recombination or CRISPR-based systems have been developed to gene editing in Lactococcus lactis, but these methods are overall inefficient. In the present study, a recombinant system based on CRISPR/Cas9 technology combined with Red/ET was developed using the plasmid pMG36e derived from Lactococcus lactis. Then, the developed recombinant system was applied to Lactococcus lactis. Knockout efficiency was significantly higher using the developed system (91%). In addition, this system showed the potential to be used as a high-throughput method for hierarchical screening. Finally, a gene-edited strain was obtained, and no antibiotics or exogenous genes were introduced using the developed gene editing system. Thus, the efficient system in lactic acid bacteria was constructed and optimized.
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| http://dx.doi.org/10.1007/s10529-023-03398-4 | DOI Listing |
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