CRISPR interference (CRISPRi) is a powerful tool to link essential and nonessential genes
to specific phenotypes and to explore their functions. Here we describe a protocol for …

Precise control of gene expression is essential for flux redistribution in metabolic pathways.
Although the CRISPR interference (CRISPRi) system can effectively repress gene …

Functional genomics is the use of systematic gene perturbation approaches to determine the
contributions of genes under conditions of interest. Although functional genomic strategies …

While genome-wide transposon mutagenesis screens have identified numerous essential
genes in the significant human pathogen Streptococcus pyogenes (group A Streptococcus …

The cell envelope of Gram-negative bacteria belonging to the Burkholderia cepacia complex
(Bcc) presents unique restrictions to antibiotic penetration. As a consequence, Bcc species …

Abstract Catalytically dead Cas9 (dCas9) is a programmable transcription factor that can be
targeted to promoters through the design of small guide RNAs (sgRNAs), where it can …

Antibiotic activity is limited by the physical construction of the Gram-negative cell envelope.
Species of the Burkholderia cepacia complex (Bcc) are known as intrinsically multidrug …

CRISPR-Cas9 has revolutionized genome engineering by allowing precise introductions of
DNA double-strand breaks (DSBs). However, genome engineering in bacteria is still a …

A significant bottleneck in the discovery of novel antibiotics is the poor understanding of the
mechanism of action as it impedes compound prioritization for further development …

With the advancement of genomic engineering and genetic modification techniques, the
uptake of computational tools to design guide RNA increased drastically. Searching for …