Portable CRISPR-Cas9^N System for Flexible Genome Engineering in Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus paracasei.

ABSTRACT: Diverse Lactobacillus strains are widely used as probiotic cultures in the dairy and dietary supplement industries, and specific strains, such as Lactobacillus acidophilus NCFM, have been engineered for the development of biotherapeutics. To expand the Lactobacillus manipulation toolbox with enhanced efficiency and ease, we present here a CRISPR (clustered regularly interspaced palindromic repeats)-SpyCas9^D10A nickase (Cas9^N)-based system for programmable engineering of L. acidophilus NCFM, a model probiotic bacterium. Successful single-plasmid delivery system was achieved with the engineered pLbCas9^N vector harboring cas9^N under the regulation of a Lactobacillus promoter and a cloning region for a custom

SUBMITTER: Goh YJ 

PROVIDER: S-EPMC8105030 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature