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. Author manuscript; available in PMC: 2017 Sep 20.
Published in final edited form as: Nat Biotechnol. 2017 Mar 20;35(5):463–474. doi: 10.1038/nbt.3834

Figure 2. 490,000-element DrugTarget-CDKO library can be efficiently assembled and delivered into cells and shows no positional bias.

Figure 2

(a) Cumulative distribution of sequencing reads for double-sgRNAs. Read counts were normalized by total reads of each sample and the cumulative sums of double-sgRNAs were plotted as relative percentages of the number of expected double-sgRNAs. (b) Histogram showing the number of double-sgRNAs per gene pair. 98.7% of the 42,319 detected gene pairs have more than 6 double-sgRNA combinations. (c) Growth (γ) phenotypes for two single genes and the corresponding gene pair were calculated from the double-sgRNA frequencies in the T14 sample and plasmid library. For γ phenotypes of single genes, all possible double-sgRNA combinations of the 3 gene-targeting sgRNAs and 79 safe-sgRNAs were measured. For the γ phenotype of a given gene pair, 9 double-sgRNA combinations were measured. Blue dotted lines mark the minimum threshold for read counts (50). (d) Minimal positional bias in the DrugTarget-CDKO library. γ phenotypes of double-sgRNAs were compared between both orientations. (e) High reproducibility of measured γ phenotypes between two experimental replicates.