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. Author manuscript; available in PMC: 2022 Apr 29.
Published in final edited form as: Cell. 2021 Apr 9;184(9):2503–2519.e17. doi: 10.1016/j.cell.2021.03.025

Figure 4. Genome-wide gene silencing by CRISPRoff.

Figure 4.

(A) A schematic of the dual sgRNA lentiviral vector used in the CRISPRoff genome-wide screens that contains two unique sgRNAs targeting the same gene.

(B) A schematic of a pooled genome-wide screen to determine the targeting landscape of CRISPRoff.

(C) A time course of CLTA expression in HEK293T after transfection of dCas9-KRAB (gray), CRISPRoff-V2 (black), or mutant CRISPRoff-D3AE765A (orange).

(D) A comparison of phenotype scores (γ) between CRISPRoff (y-axis) and CRISPRoff mutant (x-axis) screens. Three types of expected negative controls are highlighted as negative control pseudo-genes (blue), olfactory genes (orange), and Y chromosome genes (green).

(E) A violin plot of the phenotype scores (γ) for genes defined as essential or nonessential from DepMap. Each replicate screen is plotted for CRISPRoff (green) and CRISPRoff mutant (orange).

(F) A plot of true and false positive rates of genes defined as essential by DepMap.

(G) A plot illustrating the distance of an essential gene hit, defined as having a γ ≤ −0.2, from the nearest essential gene hit. Each dot corresponds to a gene hit’s nearest neighboring essential gene, with the x-axis showing the distance between the two genes and the y-axis as the neighboring gene’s phenotype score.