The DLGAP5 Knockout HAP1 Polyclonal Cells offer a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DLGAP5 gene in HAP1 cells. This loss-of-function model eliminates DLGAP5, a key mitotic spindle assembly factor, through CRISPR/Cas9-mediated gene disruption. The polyclonal format maintains genetic diversity typical of CRISPR pools, enabling robust phenotype screening and functional rescue experiments without clonal artifacts. As a genetically defined tool, it facilitates studies of mitotic mechanisms and mitotic-targeted compound evaluation in a near-haploid genomic context.
HAP1 cells are near-haploid human cells derived from the KBM-7 chronic myeloid leukemia line, displaying an adherent fibroblast-like morphology. Their haploid karyotype eliminates allelic complexity, ensuring clear genotype-phenotype correlations in knockout studies. Widely employed in functional genomics and drug sensitivity profiling, HAP1 cells preserve intact mitotic and cell cycle regulation, making them an ideal system for dissecting mitosis-specific functions through targeted gene disruptions.
DLGAP5 functions as a microtubule-associated protein essential for bipolar spindle assembly and chromosome alignment. It is recruited to spindle microtubules by TPX2 and phosphorylated by Aurora A kinase (AURKA), promoting kinesin-5 (KIF11/Eg5)-mediated spindle pole separation and stabilizing kinetochore fiber attachments. DLGAP5 interacts with key mitotic regulators including importin ??/??, survivin (BIRC5), INCENP, CDK1, and cyclin B, integrating signals from Ran GTPase and CDK1/cyclin B. This network positions DLGAP5 centrally in mitotic fidelity and spindle assembly pathways.
In the HAP1 haploid background, DLGAP5 knockout amplifies the penetrance of mitotic phenotypes, enabling clear detection of spindle defects, chromosome misalignment, and mitotic delays. This model enhances studies of synthetic lethal interactions and chemotherapeutic sensitivities, particularly relevant for cancers overexpressing DLGAP5. The simplified genome aids in differentiating DLGAP5-dependent functions from redundant spindle assembly mechanisms. Additionally, HAP1 adherence supports high-resolution live-cell imaging for detailed kinetic analyses of mitotic progression and catastrophe.
Research applications span cancer cell biology, mitosis-targeted therapy validation, and functional genomics. Key techniques include Western blotting for DLGAP5, immunofluorescence of spindle markers (??-tubulin, pericentrin, CREST), flow cytometry for DNA content, and time-lapse microscopy of aberrant mitoses. Co-immunoprecipitation with AURKA and rescue using siRNA-resistant DLGAP5 confirm target specificity, while viability assays with taxol or monastrol assess drug sensitivities. For further technical information or to discuss custom applications, please contact Ascent Research.
