Fig. 2. Genome-wide CRISPR-Cas9 screen for regulators of NOXA-induced MCL1 degradation.

a Selection strategy to enrich cells in which the ability of NOXA to provoke MCL1 degradation has been disabled. Bax^−/−Bak^−/− MEFs engineered to express GFP-MCL1 along with HA-NOXA and Cas9 (i) were transduced with a genome-wide sgRNA lentivirus library [24]. Following transduction, a small proportion of GFP^high cells was detected (ii) and enriched through three rounds of flow cytometric sorting (iii–v). b Quantitation of sgRNA abundance in the transduced cells before and after sorting. Multiple independent sgRNAs targeting Mcl1, March5, Mtch2, and Ube2k were enriched within the sorted GFP^high cell population. These data are also summarized in Supplementary Table S3.