Fig. 2. Next-generation sequencing reveals efficient and reproducible editing.

A Schematic of the deep-sequencing approach to quantify mutational efficiency for each gRNA. B Histograms depicting the allelic knockout efficiency for the most efficient (red), middle (green), and least-efficient (blue) gRNA per gene. Median values of each group are indicated by dashed vertical lines. C Representative scatterplot of mutational efficiency in a subset of randomly selected gRNA across two independent donors (r² mean = 0.745, range 0.722–0.772 over 100 randomizations). D Scatterplot of mutational efficiency versus the log₂ fold change in infection relative to the plate median for each gRNA at days 3 (left), 5 (middle), and 7 (right). Each gRNA dot is colored by phenotypic outcome based on empirically determined cutoffs to achieve a 1% false-positive rate.