Fig. 3

Impact of sgRNA features on library design. a Scatter plot showing log2 fold change of sgRNAs targeting CASP8 relative to their exon location. The gene models of major transcripts of CASP8 are depicted (ENST00000432109, ENST00000392258, ENST00000323492, ENST00000264275, ENST00000358485). b Box plots showing fold change of all sgRNAs targeting the first exon of CASP8 compared with other targeted exons. sgRNAs against the first exon are less enriched (*p ≤ 0.05, two-sided t-test). c Comparison of sgRNA features between functional and non-functional sgRNAs. All sgRNAs of BAX, FADD, and CASP8 were selected for analysis. Fold changes of individual sgRNAs of each gene were compared with the mean fold change of all sgRNAs of the respective genes. sgRNAs with a z-score >1 were grouped as functional and those with a z-score < −1 were grouped as non-functional. The on-target score by Doench et al. was calculated for both groups (y-axis) and are presented as box plots. Differences between groups were determined by a two-sided t-test. d Line graph showing interdependence between number of targetable human protein coding genes (y-axis), sgRNA coverage per gene (x-axis), and number of off-targets (colored lines). Off-targets are defined as genomic regions with at least 12 bases of homology to the protospacer of the on-target