Fig. 3. In4mer platform for whole-genome screening.

A Inzolia human whole-genome library targets single genes and paralog pairs, triples, and quads with arrays of 4 Cas12a gRNA. Each gene or gene family is targeted by two arrays encoding the same gRNA in different order. Commercially synthesized oligo pools are cloned into the one-component pRDA_550 lentiviral vector; schematic created in Biorender. B–E Screening in K-562 CML cells (prototype library), A549 lung cancer cells (prototype library) and A375, MELJUSO melanoma cancer cells (Inzolia library). B Read counts from the plasmid and experimental timepoints after lentiviral transduction. C Correlation of sample read counts. Endpoint replicates are highly correlated. D Fold change distributions of arrays targeting reference essential (red) and non-essential genes (blue) in four cell lines. E Precision/recall analysis from ranked mean fold change of arrays targeting each gene, calculated against reference essential and non-essential genes.