The KIF2C Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the loss-of-function study of KIF2C (MCAK) in a haploid human cell background. This product provides a mixed population of HAP1 cells carrying targeted disruptions in the KIF2C gene, enabling efficient investigation of KIF2C-dependent mitotic processes without clonal selection. As a polyclonal knockout resource, it captures the heterogeneity of CRISPR-induced edits, making it suitable for pooled screening and genotype-phenotype correlation studies.
The host HAP1 cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia (CML) cell line. HAP1 cells maintain a predominantly haploid karyotype, which simplifies genetic manipulation and functional genomics by reducing genetic redundancy. This unique feature makes HAP1 cells particularly valuable for knockout and mutagenesis screens, as single-allele disruption can directly reveal recessive phenotypes. The HAP1 background retains key mitotic and signaling pathways, including those governing cell cycle checkpoints and chromosome segregation, providing a physiologically relevant context for studying mitotic regulators like KIF2C.
KIF2C, also known as mitotic centromere-associated kinesin (MCAK), is a kinesin-13 family microtubule depolymerase that specifically localizes to kinetochores, spindle poles, and microtubule plus ends to regulate microtubule dynamics during mitosis. It is phosphorylated and activated by Aurora A kinase, Aurora B kinase, Plk1, and CDK1, and interacts with kinetochore complex proteins such as KNL1 and the Mis12 complex, plus-end tracking proteins EB1 and CLIP-170, and tubulin dimers. KIF2C depolymerizes microtubules to correct erroneous kinetochore-microtubule attachments and facilitate chromosome alignment and segregation. Its activity is integral to the spindle assembly checkpoint (SAC) and the G2/M transition, ensuring faithful chromosome segregation.
In the haploid HAP1 background, disruption of KIF2C is anticipated to cause severe mitotic defects, including aberrant spindle morphology, chromosome misalignment, and increased aneuploidy, modeling chromosomal instability observed in many cancers. The polyclonal nature of the knockout population allows for the observation of a range of phenotypic severities, which can be correlated with genotypic diversity. This system is particularly powerful for functional genomics screens aimed at identifying synthetic lethal interactions or modulators of the mitotic checkpoint, as the haploid state eliminates the masking effect of a second functional allele.
Typical applications include investigating cancer cell division mechanisms, validating mitotic inhibitors as therapeutic targets, and modeling chromosomal instability in drug response assays. The cells can be used in immunofluorescence to visualize mitotic spindle architecture, live-cell imaging to track chromosome movements, Western blotting to confirm KIF2C ablation and downstream signaling changes, karyotyping and aneuploidy detection, clonogenic survival assays following drug treatment, and siRNA rescue experiments to verify phenotype specificity. For further information or to inquire about custom gene-editing services, please contact Ascent Research.
