Fig. 2

Copper-inducible reporter gene activation by dCasEV2.1 in N. benthamiana. a Schematic representation of dCasEV2.1 components. b Genetic constructs for dual constitutive (Cs/Cs) transcription (35S promoter for both dCas9:EDLL and MS2:VPR), single copper-regulated (Cu/Cs) transcription (CBS:minimal DFR promoter for dCas9:EDLL and 35S promoter for MS2:VPR), and dual copper-regulated (Cu/Cu) transcription (CBS:minimal DFR promoter for both dCas9:EDLL and MS2:VPR). c Efficiency of single vs. dual copper-regulated transcription of dCasEV2.1 under a minimal DFR promoter (minDFR) to activate a pDFR:FLuc reporter. d Efficiency of dual copper-regulated transcription of dCasEV2.1 under a minimal 35S (min35S) promoter to activate a pDFR:FLuc reporter. Activation is expressed as Normalized FLuc/RLuc ratios of N. benthamiana leaves transiently expressing the genetic constructs. All FLuc/RLuc ratios were normalized using the FLuc/RLuc ratios of N. benthamiana leaves expressing a constitutive pNOS:Fluc reporter. Error bars indicate SD (n ≥ 3). Statistical analyses were performed using one-way ANOVA (Tukey’s multiple comparisons test, P-Value ≤ 0.05). Variables within the same statistical groups are marked with the same letters. The figure includes images from Biorender (biorender.com)