Fig. 4
From: Efficient prioritization of CRISPR screen hits by accounting for targeting efficiency of guide RNA
New analysis method with v metric reduces bias generated from the conventional method. A Theoretical minimum fold change as a function of indel frequency (viability = 0). B Scatter plots for indel frequency and fold change at day 21 for sgRNAs targeting indicated genes. The black line indicates the theoretical minimum FC at viability = 0. Gray shade indicates the area below theoretical minimum FC. C Quantification of cells at expected viability below 0 from B (gray). D Two theoretical scenarios showing the relationship between the reporter sequence and endogenous sgRNA target gene. E Quantification of indel frequencies of the reporter sequence and endogenous HPRT1 gene using four different gRNAs targeting HPRT1 in the presence and absence of 6-TG. F Linear regression of indel frequency and fold change at day 21 of negative control gRNAs (solid line). The dotted line is an expected line at viability = 0. Gray dots indicate individual points of negative control sgRNAs and the black bold dot indicates a hypothetical point. G The moving median of 4 genes according to X analyzed by LFC or Log2v. All data are presented as mean ± s.d. n = 3 biological replicates. Pearson correlation coefficient r is represented by R with X > 0.44 cutoff and the p value is calculated by a two-tailed test. H The receiver operating characteristic (ROC) curve of the two analysis methods. The true positive rate is decided by RPL8 and RPL15 and plotted against the false positive rate, decided by CCR5 and CD4 (left). The area under the curve analysis for essential (solid line) and non-essential (dotted line) genes (right)