The DLGAP5 Knockout HEK293T Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HEK293T cells with targeted disruption of the DLGAP5 gene. This heterogeneous pool of loss-of-function mutations enables functional analysis of mitotic spindle regulation without clonal selection bias. The CRISPR/Cas9-mediated gene disruption provides a robust model for investigating DLGAP5-dependent processes in a widely used human cell line.
HEK293T cells are adherent, epithelial-like human embryonic kidney cells immortalized with adenovirus 5 DNA and expressing SV40 large T antigen. They are widely employed for high-efficiency protein expression, viral packaging, and gene editing due to their exceptional transfectability. Their embryonic kidney origin and robust proliferation make them a relevant and tractable system for studying fundamental cell cycle and mitotic mechanisms.
DLGAP5 is a microtubule-associated protein essential for mitotic spindle assembly and stabilization, facilitating chromosome congression and cytokinesis. It is regulated by mitotic kinases CDK1, Aurora A, and PLK1, and the transcription factor FOXM1, with activity modulated by the RanGTP gradient. DLGAP5 interacts with TPX2 and Aurora A, recruits Kif11 to microtubules, and cooperates with HURP and importin ??. Its depletion disrupts spindle morphology, causes chromosome misalignment, mitotic arrest, and apoptosis, underscoring its critical role in mitotic progression.
In the HEK293T context, DLGAP5 knockout provides a valuable model for dissecting mitotic signaling networks implicated in cancer. DLGAP5 overexpression is associated with poor prognosis in hepatocellular carcinoma, breast, lung cancer, and glioblastoma. The polyclonal knockout population mimics tumor heterogeneity, enabling studies of population-level responses to DLGAP5 loss. Researchers can explore synthetic lethal interactions, evaluate mitotic vulnerability, and assess the impact of spindle defects on proliferation using this genetically tractable system.
This polyclonal knockout population is ideally suited for anti-mitotic drug screening, mitotic checkpoint studies using live-cell imaging to monitor division errors, and functional genomics screens for synthetic lethality. Representative assays include immunofluorescence to visualize aberrant spindle morphology, flow cytometry for cell cycle profiling (e.g., phospho-histone H3 staining), western blotting for mitotic regulators such as cyclin B1, and viability assays to measure apoptosis. For further details, please contact Ascent Research.
