Fig. 2
From: Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria
Optimization on synthetic UAS sites and sgRNA-dependent dynamic ranges. a Spatial relationship between the activator and the core PpspA with an artificial UAS LEB2 (Supplementary Data 1). Activator was driven by the constitutive promoter BBa_J23106. Concentrations of inducers used in the ON state: 0.4 µM AHL (sgRNA) and aTc 2.5 ng mL−1 (dCas9). No inducers were added in the OFF state. b sgRNA-dependent dynamic range optimization by “Buffer Terminators” which were previously characterized59. c sgRNA-dependent dynamic range optimization by mismatches on the sgRNA scaffold. d sgRNA-dependent dynamic range optimization by moving RNA aptamer to different positions. The first three designs contained a wild-type U-A pair at the “+5” site. The circuits for b–d all employed PpspA with two synthetic UAS LEA2 (Supplementary Data 1) and the activator was driven by the constitutive promoter BBa_J23106. Concentrations of inducers used: 0.4 µM AHL (sgRNA) and aTc 2.5 ng mL−1 (dCas9). Error bars, s.d. (n = 3); a.u., arbitrary units. Source data are provided as a Source Data file
