Figure 1: Strategies to abrogate the activity of guide RNAs by appending blocking sequences.
From: Aptazyme-embedded guide RNAs enable ligand-responsive genome editing and transcriptional activation
(a) Schematic representation of the essential regions of the guide RNA that were targeted by candidate blocking sequences. (b) In vitro activity of unmodified guide RNAs and three blocked guide RNA variants depicted in the lower right. The canonical and blocked sgRNAs were each incubated with the target dsDNA (the full-length GFP gene) in the presence of Cas9 for 1 h at 37 °C, while the crRNA and tracrRNA variants were incubated for an additional 4 h due to the lower overall activities of the corresponding Cas9:guide RNA complexes.
