Electroporation of Cas9 RNP by Neon Transfection system. (A) We screened 41 pulse settings to electroporate CD96-targeting Cas9 RNP. Condition 7 is a reference condition reported by Pomeroy et al. (2020) for cas9 mRNA. (B) Conditions 7, 8, and 9 were examined in triplicate. Cell viability and viable CD96⁻ cells were determined as described for the Lonza system. The data are shown as mean ± SD of three donors (n = 3). Two-tailed Welch’s unequal variances t test was used to test for statistical significance. *, P ≤ 0.05; **, P ≤ 0.01. Std, standard for comparison.