Cloning of individual sgRNAs and sgRNA libraries

For the 6K-cancer library, we used the lentiCRISPR v2 vector (plasmid No. 52961, Addgene, Watertown, MA) as backbone [56]. We designed ten sgRNAs per gene to target ∼6000 genes and added non-targeting sgRNAs as controls (Table S1). For library construction, we used the same protocol as previous CRIPSR screen [52]. For individual sgRNA cloning, we synthesized the pairs of oligonucleotides (IDT) containing the BsmBI-compatible overhangs. We used the standard protocols to anneal and clone the sgRNA oligos into the lentiCRISPR v2 vector [56]. The sequences of individual sgRNAs for CDK6 and ETV5 are shown in Table S8.