Implementation of the Argonaute Genome Editing Protein in Lactococcus Lactis

Probiotics are microorganisms that, when administered in adequate amounts, confer a health benefit on the host. For example, Lactococcus lactis (L. lactis), commonly found in dairy products, alleviates inflammatory bowel disease and has been engineered to excrete therapeutic proteins. However, the tools to genetically manipulate L. lactis remain limited and low-throughput. We hypothesize that the Natronobacterium gregoryi Argonaute protein (NgAgo) can be used as a genome editing method in L. lactis. Recently demonstrated to effectively promote genetic recombination in mammalian cells, NgAgo employs a complementary single-stranded DNA as a guide to cleave genomic DNA. By incorporating the NgAgo gene into L. lactis, we test the system to select for mutations at exact sights within the genome. Our work should result in a genome editing tool that is more selective and easier to use than the current industry standard, CRISPR/Cas9, with potential downstream applications in all-DNA-based rapid multiplexing technologies.