Figure 5: Experiments validate that high sensitivity is maintained at low cell counts and high MOI.

A) CRISPR FACS screens to identify regulators of IL2RA were performed at low (~0.3) and high (~2) MOI and low and high coverage. Low coverage (average of 195x coverage for MOI 0.3, 208x coverage for MOI 2) and high coverage (average of 1662x coverage for MOI 0.3, 1180 x coverage for MOI 2) n = 3 donors. The relative number of cells collected for each condition is shown. B) Quantification of the number of viral integrations per cell using droplet digital PCR in the unsorted cells (total) or after sorting cells that express GFP in the gRNA lentiviral construct (GFP+). C) Comparison of screens hits from the high coverage, low MOI screen vs low coverage, high MOI screen analyzed using Waterbear. D) Experimentally validated regulators of IL2RA were detected as screen hits in the low coverage, high MOI screen using Waterbear.