Fig. 1: Arrdc3 identified as a hit in a CRISPR/Cas9 screen for regulators of TRP53-mediated lymphoma growth suppression.
From: Genome-wide CRISPR screening identifies a role for ARRDC3 in TRP53-mediated responses
A Diagram of the CRISPR screen methodology using the MDM2 inhibitor nutlin-3a, which activates TP53/TRP53 in a non-genotoxic manner, as a selection pressure. In quadruplicate, AF47A Eμ-Myc lymphoma cells were transduced with the whole-genome “Yusa” sgRNA library, expanded, and either cultured without treatment or treated with either DMSO (vehicle control) or nutlin-3a (10 μM) for 24 h. Live cells were then sorted by FACS, DNA extracted from each sample (marked by asterisks), indexing PCR performed, and NGS combined with bioinformatic analyses used to identify sgRNAs that conferred a survival/competitive advantage after TRP53 activation. B Top hits from the CRISPR screen, comparing the untreated control cells (yellow asterisk) with the cells treated with nutlin-3a (red asterisk). Arrdc3 was identified as one of the top hits, with Arrdc3 targeting sgRNAs highly enriched in the surviving nutlin-3a-treated lymphoma cells.
