Pooled in vitro and in vivo CRISPR-Cas9 screening identifies tumor suppressors in human colon organoids

UMI validation screen, original data from in vivo tumor suppressor CRISPR/Cas9 screening experiment. Figure 6.
Experimental setup: 281 tumor suppressor gRNAs, positive controls, neutral controls and gRNAs targeting essential genes were introduced into the CRISPR-UMI lentivirus (Michlits et al., 2017). Transduced AK organoids (APC-KO and KRASG12D) were transplanted subcutaneously in NSG mice followed by barcode sequencing of the tumor pool (from 10 mice) and 5 individual tumors. Clone and read number were determined by next generation sequencing. The conventional gRNA enrichment analysis was compared to UMI-CRISPR-screening with outlier removal.