GSM8783906: T0, biological replicate 2; Homo sapiens; OTHER
gDNA from all cells was harvested and sgRNA barcodes were amplified and processed for sgRNA abundance library preparation using Q5 High-Fidelity DNA Polymerase (NEB) and sequenced on an Illumina NovaSeq 6000 We used a previously optimized and validated, compact, highly active sgRNA library containing the top 2 on-target sgRNAs for 23,483 genes cloned into the library expression vector pU6-sgRNA EF1alpha Puro-T2A-GFP derived from pJR85(#140095, Addgene) and modified to express a second sgRNA using the human U6 promoter as previously described. Knockdown efficiency of all guide sequences is publicly available at https://gwps.wi.mit.edu. 1137 non-targeting sgRNA pairs were also included as negative controls in the screen.