Figure 3
From: HideRNAs protect against CRISPR-Cas9 re-cutting after successful single base-pair gene editing
Parameters affecting hRNA performance. (A) Three hRNAs using different PAMs (1, 2 and 3, marked blue, red and green, respectively) and varying protospacer lengths were designed to protect the activated GFP reporter against re-cutting. Part of the GFP reporter sequence after repair with the ATG only template is shown (T of ATG marked green) and the PAMs. Part of the spacer sequence of the hRNAs that target PAMs 1–3 is shown, with the A that marks their specificity marked in green. (B) Protection scores of hRNAs 1–3 with different spacer lengths determined at three different puromycin concentrations. If the first nucleotide of the protospacer was not a G, a G was added to the sequence. The hRNA protospacer length does not take a mismatching 5′ G into account. Error bars denote standard error of mean (n = 3).
