title: Occurrence and activity of a type II CRISPR-Cas system in Lactobacillus gasseri.
abstract: Bacteria encode clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated genes (cas), which collectively form an RNA-guided adaptive immune system against invasive genetic elements. In silico surveys have revealed that lactic acid bacteria harbour a prolific and diverse set of CRISPR-Cas systems. Thus, the natural evolutionary role of CRISPR-Cas systems may be investigated in these ecologically, industrially, scientifically and medically important microbes. In this study, 17 Lactobacillus gasseri strains were investigated and 6 harboured a type II-A CRISPR-Cas system, with considerable diversity in array size and spacer content. Several of the spacers showed similarity to phage and plasmid sequences, which are typical targets of CRISPR-Cas immune systems. Aligning the protospacers facilitated inference of the protospacer adjacent motif sequence, determined to be 5'-NTAA-3' flanking the 3' end of the protospacer. The system in L. gasseri JV-V03 and NCK 1342 interfered with transforming plasmids containing sequences matching the most recently acquired CRISPR spacers in each strain. We report the distribution and function of a native type II-A CRISPR-Cas system in the commensal species L. gasseri. Collectively, these results open avenues for applications for bacteriophage protection and genome modification in L. gasseri, and contribute to the fundamental understanding of CRISPR-Cas systems in bacteria.
authors: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors: Rosemary
authors: R
authors: Sanozky-Dawes
authors: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.|Functional Genomics Program, North Carolina State University, Raleigh, NC, USA.
authors: Kurt
authors: K
authors: Selle
authors: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors: Sarah
authors: S
authors: O'Flaherty
authors: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.|Functional Genomics Program, North Carolina State University, Raleigh, NC, USA.
authors: Todd
authors: T
authors: Klaenhammer
authors: Functional Genomics Program, North Carolina State University, Raleigh, NC, USA.|Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA.
authors: Rodolphe
authors: R
authors: Barrangou
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D023061:Gene Order; D016680:Genome, Bacterial; D007778:Lactobacillus / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics