A genome-wide CRISPR-activation screen with Eg5 inhibition in haploid cells
Genome-wide CRISPR-activation screening was performed in human somatic haploid cells to identify genes that promote the maintenance of the haploid state and improve chromosome stability.Prior to the screen, we found that the human somatic haploid cells exhibited attenuated mitotic spindle bipolarization in an Eg5-dependent manner. Pharmacological inhibition of Eg5 accelerated chromosome instability in haploid cells. Supplementing with Cep192, a centrosomal scaffold protein, restored spindle bipolarization and chromosome stability to diploid levels under Eg5 inhibition. We expanded our approach to a genome-wide scale in the pooled CRISPR-activation screen and identified 133 candidate genes that could promote chromosome stability in haploid cells.