Cas9 off-target binding to the promoter of bacterial genes leads to silencing and toxicity.

Rostain W, Grebert T, Vyhovskyi D, Pizarro PT, Tshinsele-Van Bellingen G, Cui L, Bikard D
Nucleic Acids Res. 2023 Mar 17:gkad170. doi: 10.1093/nar/gkad170. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
198639	pLC133	Expression plasmid for dCas9 in E. coli used in Chip-seq experiments. dCas9 is tagged with a C-terminal 3x FLAG tag. A guide RNA can be cloned using BsaI restriction sites.
198640	pTG131	Thermosensitive pSC101-based plasmid expressing a sgRNA under the control of the J23119 promoter and Cas9 under the control of the DAPG-inducible PhlF promoter