GSM6430038: A375_CRISPRi_day21_replicate1; Homo sapiens; OTHER
The sgRNA sequences were synthesized by Synbio Technologies and cloned into lentiGuide-Puro vector (Addgene, 52963) in the form of homologous recombination to construct the plasmid library, and then packaged into the lentivirus library for subsequent screening. We generated A375 cell lines stably expressing Lenti-dCas9-KRAB (Addgene, 89567) transgenes and performed lentiviral transduction of pooled sgRNAs at low multiplicity of infection (MOI) = 0.2. After 24 hours, puromycin was added for selecting the effective infected A375-KRAB cells. After 7 days, the cells were divided into two groups. One group (day 0) of cells was collected and frozen as a control group, and the other group (day 21) of cells were selected at 21 days. CRISPR interference (CRISPRi) screen on A375 enhancers