The KLHL13 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the KLHL13 gene, providing a loss-of-function model for studying its role as a substrate adaptor for CUL3-based ubiquitin ligases. This polyclonal format avoids clonal selection artifacts while maintaining effective gene disruption.
HAP1 is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells, offering simplified genetic analysis due to its single allele per gene. Its adherent growth and retention of cancer-relevant pathways make it ideal for generating knockout models to investigate cell cycle and mitotic control.
KLHL13 recruits targets such as Aurora B kinase to the CUL3-RBX1 E3 ligase complex for ubiquitination and proteasomal degradation, thereby regulating mitotic progression and cytokinesis. It interacts with CUL3, RBX1, and components of the chromosomal passenger complex (INCENP, Survivin, Borealin), and its activity is controlled by cell cycle cues and feedback from Aurora B.
In the haploid HAP1 background, KLHL13 disruption yields unambiguous loss-of-function phenotypes, enabling clear attribution of mitotic defects such as chromosome misalignment, spindle abnormalities, and cytokinesis failure. This model allows direct investigation of ubiquitin-dependent control of mitosis without confounding genetic compensation.
Typical applications include western blotting to monitor Aurora B levels, flow cytometry for cell cycle distribution, immunofluorescence for mitotic spindle assessment, and ubiquitination assays to measure substrate modification. Co-immunoprecipitation can probe CUL3 complex interactions, and time-lapse imaging can capture cytokinesis dynamics. These cells support research in cancer biology, cell division, and proteostasis. For technical inquiries, contact Ascent Research.
