Validating condition-specific GIs detected in pooled screen. (A) Heatmap depicting GI scores derived from pooled fitness assay in each of five growth conditions for 17 guide pairs. Red text indicates strains reconstructed for validation, and asterisks signify absolute z-score greater than two. (B) Positive GIs for COG8 and YCR016W with CDC25 and negative GIs for RPN5 with LSM2 and LSM4 are validated on spot assay plates in the presence of CRISPRi-inducing agent ATc. (ATc) Anhydrotetracycline, (NT) nontargeting control gRNA. (C) Positive GIs validated in monoculture for COG8 and YCR016W with CDC25 and CYR1. Error bars, 95% confidence intervals around mean. Horizontal line is 95% confidence interval around expected double-mutant fitness (using multiplicative model). Asterisks represent values passing significance based on nonoverlapping 95% confidence intervals of observed and expected double-mutant fitness.
