Fig. 2

Electroporation leads to more effective transfection and HPRT editing efficiency compared to lipid nanoparticle delivery. Cell viability, transfection and editing efficiencies were calculated upon Cas9-RNP delivery with A the lipid nanoparticles Lipofectamine 3000™, Lipofectamine™ RNAiMAX and FuGENE® or the B Neon™ electroporation system, by testing different ranges of voltage, milliseconds or applied pulses, respectively. C Neon™ electroporation program optimization, calculating % of live cells, % of GFP⁺ cells and T7E1 efficiency. Non-transfected cells were used as viability controls, while cells transfected with an incomplete RNP, consisting of a Cas9-GFP without gRNA, were used as transfection and editing efficiency controls. Data are represented as mean ± standard deviation of 3 technical replicates from one donor (n = 3). Statistical significance was determined using one-way ANOVA with a Tukey’s multiple comparisons correction (* p < 0.05, ** p < 0.01, and *** p < 0.001)