Figure 4.

Activation of coding and noncoding genes by dCas9-VP64 and multiplexed gRNAs. A, Diagram showing the intron-containing GUS reporter gene is under a synthetic minimal promoter containing all three target sites from the promoter of miR319. B, Activation of the GUS reporter gene by pco-dCas9-VP64 coupled with three gRNAs that target the synthetic promoter. C and D, These two diagrams depict gene structure and target sites of gRNAs for AtPAP1 and miR319, respectively. E and F, qRT-PCR analysis of target gene transcript levels. For each experiment, three independent transgenic lines were randomly chosen for analysis. Transgenic plants that express Cas9 nuclease with a similar T-DNA vector backbone were used as negative control (–). Error bars represent standard deviations of technical replicates (n = 3). UTR, Untranslated region.