Figure 2
From: CRISPR interference to interrogate genes that control biofilm formation in Pseudomonas fluorescens
CRISPRi silencing of genes involved in cell division and cell morphology in P. fluorescens. Cells harboring the pPFL-dCas9 plasmid and the pPFL-gRNA plasmid expressing a gRNANT targeting the start of the ORFs to block transcription elongation. Control with no guide (left panel); silencing of ftsZ (PFLU0952) using a ftsZNT guide (center panel), and silencing of mreB (PFLU0863) using a mreBNT guide (right panel). Strains were grown 5 hours in the presence or absence of inducer (aTc 0.1 μg/ml). Cells were stained with the membrane fluorescent dye FM 4–64 prior to observation by epifluorescence microscopy. Scale bars indicate 2 μm.
