Next-Generation Sequencing of sgRNA spacer sequences from a CRISPRi library screen of the Salmonella enterica serovar Typhimurium 14028s genome in response to repeated monoclonal IgA treatment

ABSTRACT: For this study, we implemented the anti-lipopolysaccharide monoclonal IgA Sal4 in a genome-wide CRISPR interference (CRISPRi) screen to identify genes that modulate the extent of agglutination of Salmonella enterica serovar Typhimurium (STm) in response to antibody exposure. We leveraged our recently-developed \"snow globe assay\" method to enrich for cells containing sgRNAs that silence the expression of genes that contribute to the process of antibody-mediated agglutination. A high density (~10^8 CFU/mL) culture of an STm strain containing a library of 36,651 sgRNA-encoded plasmids was treated with Sal4 IgA (15 μg/mL) in PBS for five hours at room temperature. The remaining portion of the culture present at the top of the supernatant after antibody treatment was passaged and grown overni

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Salmonella enterica subsp. enterica serovar Typhimurium

SUBMITTER:  

PROVIDER: E-MTAB-14834 | biostudies-arrayexpress |

SECONDARY ACCESSION(S): ERP169222

REPOSITORIES: biostudies-arrayexpress