480527 An Improved Method to Edit Undomesticated Lactobacilli Strains Using Crispr-Cas9 Systems

Monday, November 14, 2016
Grand Ballroom B (Hilton San Francisco Union Square)
Malay Shah, North Carolina State University, Cary, NC

The rise in new genetic tools have given scientists powerful ways to modify and study prokaryotes. However, many of these methods require a high transformation efficiency of the bacterial strains in order to work, which significantly limits use of these methods across species. Here, we developed a novel method utilizing CRISPR-Cas9 and repair templates to genetically edit low transformation efficient strain of abundant Lactobacillus Plantarum. By using CRISPR-Cas9 in these strains, we show that single point mutations are almost impossible to achieve without using a double stranded DNA repair template. We plan to use this method to further probe and study the diverse lactobacilli genus. This method offers a powerful means of editing lactobacilli and should be translatable to other strains of difficult to work with bacteria.

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