Gene expression of DREAM complex-related inducible knockouts in HeLa cells

The DREAM complex and interacting proteins (e.g., BMYB and FOXM1) together control cell cycle-dependent gene expression. We identified an unexpected, specific kinetochore defect phenotype after depletion of a DREAM complex member (LIN52) in optical pooled CRISPR knockout screens of essential genes. Here we generated individual doxycycline-inducible Cas9 cell lines expressing lentiviral-integrated sgRNAs targeting LIN52, MYBL2, and FOXM1 to evaluate differential roles in gene expression regulaion. Biological replicates were defined as replicate doxycycline inductions of Cas9 expression. A negative control sgRNA sequence was used that targets a single locus. N=3 biological replicates were performed for each gene target and for the negative control sgRNA (12 total samples).