Figure 4
From: HideRNAs protect against CRISPR-Cas9 re-cutting after successful single base-pair gene editing
hRNAs protecting a mutation outside the target site. (A) Top: sequences of the reporter before (WT) and after ssODN-directed insertion of TGG (ATG) or TGG insertion plus PAM disruption (ATG + PAM). The ‘ATG’ template leaves the entire Cas9 target site intact and thus sensitive to re-cutting. Bottom: gRNA and hRNA A and B spacers are shown, interacting with the target sequence after TGG insertion. (B) HideRNAs can protect a mutation that is outside of the Cas9 target site. The percentages of GFP-expressing cells are scored after puromycin selection at 3 different concentrations for combinations of Cas9 and the gRNA, the ATG, ATG + PAM ssODN template and hRNA (A) or (B). Error bars denote standard error of mean (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (Welch one-sided test, comparison with the ATG ssODN template).
