Gene expression of ZNF335, RNPC3, CLP1 and LIN52 inducible knockouts in HeLa cells

From our pooled optical screen, we identified an unexpected role for ZNF335, a transcription factor, in spindle bipolarity. We also found that RNPC3, CLP1 or LIN52 are regulators of gene expression on various levels, displayed an unexpected, specific kinetochore defect. Here we generated individual doxycycline-inducible Cas9 cell lines expressing lentiviral-integrated sgRNAs targeting ZNF335, LIN52, CLP1, and RNPC3 to evaluate differential roles in gene expression regulation. Here we generated individual doxycycline-inducible Cas9 cell lines expressing lentiviral-integrated sgRNAs targeting ZNF335, LIN52, CLP1, and RNPC3 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=2 biological replicates were performed for each gene target.