Combinatorial CRISPR loss-of-function screen for deubiquitinases and genes related to the ubiquitin proteasome system in HCT116 cells
The goal of the project is to identify deubiquitinases (DUBs) networks that could be targetable in cancer. To identify functionally interacting DUBs, we performed a pooled combinatorial knockout screen using a CRISPR/Cas12a system. Here we transduced a custom guide library targeting any 2-gene combination derived from 160 genes (including ~90 DUBs and other genes of interest) into Cas12a-expressing HCT116 cells. While the cells are subjected to puromycin selection, a reference sample (Day 4 post-transduction) was taken and sequenced. Two replicates were derived from this pool that were expanded and sequenced at Day 11 and Day 18 post-transduction.
The goal of the project is to identify deubiquitinases (DUBs) networks that could be targetable in cancer. To identify functionally interacting DUBs, we performed a pooled combinatorial knockout screen using a CRISPR/Cas12a system. Here we transduced a custom guide library targeting any 2-gene combination derived from 160 genes (including ~90 DUBs and other genes of interest) into Cas12a-expressing HCT116 cells. While the cells are subjected to puromycin selection, a reference sample (Day 4 post-transduction) was taken and sequenced. Two replicates were derived from this pool that were expanded and sequenced at Day 11 and Day 18 post-transduction.
European Bioinformatics Institute