CRISPR screen for regulators of cadherin based cell to cell adhesion

Pooled Crispr based genome wide screen to find potential (transcriptional) regulators and pathways underlying the regulation of cadherin based cell to cell adhesion. Overall design: A spontaneously immortalized human keratinocyte line (HaCaT) expressing Cas9 was transduced with lentivirus containing the Brunello library encoding for 76441 sgRNAs inclusive 1000 non-targeting controls at a coverage of approximately 400 cells/sgRNA. The transduced cells were selected and expanded maintaining a library coverage of approximately 600 cells/sgRNA. 50-60x10^6 cells were stained Desmoglein 3 (Dsg3), and the 10 % cells with the lowest and the 10 % cells with the highest expression of Dsg3, respectively, were isolated by FACS. 50-60 x10^6 cells served as a control before FACS. Two technical replicates were done (repl 1 and 2).