GSM8219648: Stable-cell, Bottom 0-2.5% (GFP Low), rep 2; Homo sapiens; OTHER
HCT116 cells were transduced by CRISPR lentivirus generated from Brunello sgRNA pooled library (Addgene #73179) and transduced cells were selected by puromycin. After puromycin selection, LNP-formulated EGFP-encoding mRNA with or without base modification was transfected to pooled KO cells, and cells with different GFP intensity was FACS sorted for subsequent library preparation. sgRNA sequence-containing region of gDNA from FACS sorted cells were PCR-amplified with TruSeq adaptor, purified using SPRI beads, then sequenced