Genome-wide CRISPR-dCas9 screens in E. coli identify essential genes and phage host factors.

ABSTRACT: High-throughput genetic screens are powerful methods to identify genes linked to a given phenotype. The catalytic null mutant of the Cas9 RNA-guided nuclease (dCas9) can be conveniently used to silence genes of interest in a method also known as CRISPRi. Here, we report a genome-wide CRISPR-dCas9 screen using a starting pool of ~ 92,000 sgRNAs which target random positions in the chromosome of E. coli. To benchmark our method, we first investigate its utility to predict gene essentiality in the genome of E. coli during growth in rich medium. We could identify 79% of the genes previously reported as essential and demonstrate the non-essentiality of some genes annotated as essential. In addition, we took advantage of the intermediate repression levels obtained when targeting the template str

SUBMITTER: Rousset F 

PROVIDER: S-EPMC6242692 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature