Figure 5. Validation of predicted genetic interactions using individual sgRNAs.
(a) Reproducibility between γ phenotypes in the primary and batch retest DrugTarget-CDKO screens. Data represent mean ± SEM. (b) Reproducibility between Norm-GIs in the primary and batch retest screens. Data represent mean ± SEM. The Pearson correlation after same-gene targeting pairs were removed is reported in parentheses. (c) Schematic for dual sgRNA validation assay. Cas9-expressing K562 cells were infected with lentiviruses expressing one sgRNA in a GFP vector and another sgRNA in an mCherry vector. The abundances of the four resulting fluorescent populations (uninfected, GFP, mCherry, and GFP+mCherry) were measured after 7 days. (d) K562 cells expressing a PIM1-targeting sgRNA (GFP) and a PIM2-targeting sgRNA (mCherry). The growth phenotype is calculated by measuring the relative depletion of the single-infected and double-infected cells at day 7 vs. day 0. (e,f) Quantification of growth phenotypes and genetic interaction scores for indicated double sgRNA infections compared to single sgRNA_safe-targeting controls (see methods). The PIM1_PIM2 sgRNA pair is synergistic as predicted while the BSG_GPI pair is buffering as predicted. Data represent mean ± SD from 3 replicate cultures. (g,h) Additional high-confidence synergistic and buffering gene pairs validate with predicted genetic interactions. Data represent mean ± SD (n=3) from replicate cultures.