Fig. 3: Comparison of editing rates with evolved PE variants in mammalian cell lines.
From: Enhancing prime editor activity by directed protein evolution in yeast
a Correct percentage of substitutions assessed by deep sequencing in cells transfected with plasmids encoding for PEmax, PE_Y17, PE_Y18 together with plasmids encoding for the enhanced prime editor guide RNAs (epegRNAs) at different time points on site 1 in HEK293T cells; not significant (ns) P = 0.5476 ****P < 0.0001, ***P = 0.0002, ****P < 0.0001, ***P = 0.0002 and **P < 0.0012 (left to right). b Editing rates of PE_Y17, PE_Y18 and PEmax at other loci in HEK293T cells, analyzed 48 h after plasmid transfection; not significant (ns) P = 0.9462, ns P = 0.2578, **P = 0.0084, ****P < 0.0001, ns P = 0.8254, *P = 0.0478, **P = 0.001, ****P < 0.0001, *P = 0.0341, ***P = 0.0008, ns P = 0.6543, ns P = 0.1331, ns P = 0.1621, ***P = 0.0002, *P = 0.011, **P = 0.0086, ns P = 0.6507, ns P = 0.6442, ns P = 0.7257 and ns P = 0.1566 (left to right). c Editing rates of PEmax, PE_Y17 and PE_Y18 on endogenous loci in K562 cells, analyzed 120 h after transfection; not significant (ns) P = 0.3622, ****P < 0.0001, ns P = 0.8171, ns P = 0.2033, **P = 0.0049, ****P < 0.0001, **P = 0.0022 and ****P < 0.0001 (left to right). d Editing rates of PEmax, PE_Y17 and PE_Y18 on a self-targeting library in K562 cells. The self-targeting library encoding for epegRNAs and their respective target sites was integrated into cells using lentiviral vectors prior to PE plasmid transfection and analysis of editing rates by deep sequencing after 120 h; not significant (ns) P = 0.305 and *P < 0.0486 (left to right). e Editing rates with PE variants encoded on mRNA and nucleofected into HEK293T cell line expressing an epegRNA targeting site 12; not significant (ns) P = 0.1152 and *P = 0.0145 (left to right). f Nucleofection of PEmax, PE_Y17 or PE_Y18 protein into HEK293T cells expressing an epegRNA targeting site 12; not significant (ns) P = 0.6306 and **P = 0.0042 (left to right). g Nucleofection of PE RNPs with a chemically modified pegRNA targeting site 4 in HEK293T cells; **P = 0.0079, ***P = 0.0002 (left to right). Data are displayed as means±s.d. of three independent experiments (n = 3) and were analyzed using a two-way ANOVA using Tukey’s multiple comparisons.
