The KIF20B Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the functional disruption of the KIF20B gene in a near-haploid human cell background. This product provides a heterogeneous pool of edited cells harboring targeted gene disruptions, enabling loss-of-function studies without clonal selection. The polyclonal format preserves population-level diversity while avoiding artifacts that can arise from clonal isolation, making it suitable for pooled screening and robust phenotypic analysis. Researchers can directly employ these cells to investigate the role of KIF20B in cell division and genomic stability, leveraging the genetic tractability of the HAP1 host line.
The host cell line HAP1 is a near-haploid fibroblast-like cell population derived from the KBM-7 chronic myeloid leukemia lineage. Its haploid karyotype simplifies genetic manipulation and facilitates phenotype-to-genotype correlations, making it a powerful platform for genetic screens, drug target validation, and functional genomics. HAP1 cells retain key mitotic and cytokinetic machineries, and their adherent growth and stable chromosome content under culture conditions enable reproducible quantitative assays. The near-haploid state amplifies the consequences of gene loss, generating clear phenotypic readouts for genes such as KIF20B that influence ploidy maintenance and cell cycle progression.
KIF20B encodes a kinesin-6 motor protein that functions as a critical regulator of abscission, the final step of cytokinesis. It localizes to the central spindle and midbody, where it promotes microtubule bundling and facilitates the recruitment of the ESCRT-III complex, including VPS4 and TSG101, to sever the intercellular bridge. KIF20B is phosphorylated by and interacts with PLK1 and Aurora B, and is recruited in cooperation with PRC1 and MKLP1. Its activity is tightly coordinated with the cell cycle; upstream activation by the CDK1/Cyclin B complex ensures timely engagement. Disruption of KIF20B leads to cytokinesis failure, evidenced by the arrest of cells in a binucleated state, and consequently contributes to genomic instability.
In the HAP1 background, disruption of KIF20B generates a clear and quantifiable cytokinesis defect, making this polyclonal knockout population a relevant model for investigating the molecular basis of abscission failure and its consequences for genome integrity. The near-haploid state sensitizes cells to ploidy shifts, allowing rapid detection of binucleation by flow cytometry or microscopy. This model is particularly valuable for studying the pathways that link mitotic exit to abscission, and for screening factors that cooperate with or bypass KIF20B function, thereby addressing mechanisms underlying aneuploidy and cancer-associated chromosomal instability in diseases such as breast, lung, and hepatocellular carcinoma.
These knockout cells are well suited for a range of hypothesis-driven and screening-based applications. Key experimental approaches include immunofluorescence microscopy to assess midbody formation and ESCRT-III localization, Western blotting to confirm KIF20B protein ablation, flow cytometry for ploidy and cell cycle profiling, time-lapse imaging to track cytokinesis dynamics, and cell proliferation assays. The polyclonal population can be used to identify synthetic lethal interactions, screen chemical modulators of abscission, and dissect the interplay between KIF20B and its interacting partners such as PLK1, Aurora B, and PRC1. For additional details or to discuss custom applications, please contact Ascent Research.
