Mobile-CRISPRi as a tool for genetic manipulation in the intracellular pathogen <i>Piscirickettsia salmonis</i>.

ABSTRACT: Piscirickettsia salmonis is the causative agent of salmonid rickettsial septicemia (SRS), the main bacterial disease affecting the salmon industry in Chile. In this work, we implemented a Mobile-CRISPRi system to generate gene silencing using a catalytically inactive dCas9 protein and an isopropyl β-D-1-thiogalactopyranoside (IPTG)-inducible single-guide RNA (sgRNA). We demonstrate the efficacy of the CRISPRi system in P. salmonis by silencing an exogenous reporter (sfGFP) and an endogenous regulator (Fur) that controls intracellular iron homeostasis in bacteria. The inducible expression of dCas9 and the sfGFP-directed sgRNA caused a 98.7% decrease in fluorescence in the knockdown strain. This silencing system was effective in seven P. salmonis strains fr

SUBMITTER: Ortiz-Severin J 

PROVIDER: S-EPMC12838400 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature