Figure 3: Small molecule-controlled genome editing and transcriptional activation enabled by agRNAs in human cells.

(a) Schematic representation of the activation of theophylline-agRNA in the presence of theophylline. (b) Genome-editing activity of theophylline-agRNA in HEK293-GFP cells. Endogenous GFP sites were edited by the Cas9:guide RNA complex upon agRNA activation. Cells that were not transfected with any guide RNA plasmid served as the negative control, while cells transfected with Cas9 and a canonical sgRNA (+) served as a positive control. The GFP fluorescence loss was quantified by comparing the mean cell fluorescence in transfected cells to that in cells treated with lipids only. (c) Schematic representation of the activation of guanine-agRNA in the presence of guanine. (d) Transcriptional activation activity of guanine-agRNA in the presence and absence of the ligand in HEK293T cells. GFP activation was achieved using dCas9-VPR:guide RNA complex upon agRNA activation. Values and error bars reflect mean GFP fluorescence and the s.d. of three biological replicates.