Transcription activation via PaeCascade-RpoD in S. oneidensis MR-1
(A) The schematic diagram and mechanism of PaeCascade-RpoD to recruit RNA polymerase (RNAp). RpoD (σ70) binds to transcriptional regulatory region and recruits the core RNAp (ɑ2ββ′ω) to enable gene activation. RpoD is fused to Cas7, responsible for binding protospacer.
(B) Location sites of designed crRNAs targeting TSS upstream of gfp in the plasmid pTarget.
(C) Plasmid map of PaeCascade-RpoD for transcriptional activation. The plasmid pPaeR is consisted of Cascade fused with RpoD proteins and a designed crRNA cassette controlled by Ptac and PCI promoter, respectively.
(D) The activation efficiency of type I-F PaeCascade-RpoD system targeting gfp. The values above the bar indicate the activation folds. Median GFP levels are normalized to the cr0 control strain. Values and error bars indicate mean ± SEM of three replicates.
See also Figure S1.