Morphological and Transcriptional Responses to CRISPRi Knockdown of Essential Genes in Escherichia coli.

ABSTRACT: CRISPR interference (CRISPRi) has facilitated the study of essential genes in diverse organisms using both high-throughput and targeted approaches. Despite the promise of this technique, no comprehensive arrayed CRISPRi library targeting essential genes exists for the model bacterium Escherichia coli, or for any Gram-negative species. Here, we built and characterized such a library. Each of the ∼500 strains in our E. coli library contains an inducible, chromosomally integrated single guide RNA (sgRNA) targeting an essential (or selected nonessential) gene and can be mated with a pseudo-Hfr donor strain carrying a dcas9 cassette to create a CRISPRi knockdown strain. Using this system, we built an arrayed library of CRISPRi strains and performed population and single-cell growth and m

SUBMITTER: Silvis MR 

PROVIDER: S-EPMC8510551 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature