Figure 1.

General workflow of CRISPR screens. CRISPR screening protocols include 6 main steps: (1) in silico design of gRNA libraries; (2) cloning and validation of gRNA library; (3) packaging gRNAs into lentiviruses and transduction of lentiviruses into Cas-expressing cells; (4) alteration of genes in experimental conditions; (5) isolation of nucleic acids; and (6) deep sequencing of barcoded gRNAs and data analysis. Note: if using pre-made gRNA libraries, the first two steps of the protocol are omitted.