DYRK4 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cervical adenocarcinoma cell line, engineered to disrupt the dual-specificity tyrosine-phosphorylation-regulated kinase 4 (DYRK4) gene. This knockout model serves as a robust experimental tool for dissecting the role of DYRK4 in cytokinesis and cell cycle progression. The polyclonal format provides a heterogeneous knockout pool that reflects the variability of CRISPR-mediated gene disruption, enabling functional studies without the need for single-cell cloning. Loss of DYRK4 function in these cells allows researchers to interrogate its kinase activity and downstream targets in a human cellular context relevant to cancer biology.
The host cell line, HeLa, is an immortalized human cervical adenocarcinoma cell line widely utilized in biomedical research due to its robust growth characteristics and amenability to genetic manipulation. Derived from a patient with cervical cancer, HeLa cells are positive for human papillomavirus type 18 (HPV-18) and exhibit an epithelial morphology. This background provides a well-characterized platform for studying oncogenic processes and mitotic regulation. HeLa cells are particularly suited for high-resolution imaging applications due to their flat, adherent growth, facilitating detailed analysis of subcellular structures such as the mitotic spindle and midbody.
DYRK4 encodes a dual-specificity protein kinase that undergoes autophosphorylation on tyrosine residues and accumulates at the spindle midzone and midbody during cytokinesis. It phosphorylates septin family proteins, including SEPT2 and SEPT7, promoting their filament organization and contributing to contractile ring constriction. DYRK4 functions downstream of mitotic kinases such as CDK1 and PLK1, and it interacts with 14-3-3 proteins and the centralspindlin complex components MKLP1 and MGCRacGAP. Representative pathway components include Aurora B kinase, PLK1, the centralspindlin complex, septin cytoskeleton, and the actomyosin contractile ring. Disruption of DYRK4 impairs septin phosphorylation, leading to defective cytokinetic furrow assembly.
In the HeLa cellular context, knockout of DYRK4 results in cytokinesis failure, manifesting as the accumulation of binucleated and multinucleated cells. This phenotype is a direct consequence of disrupted septin dynamics and actomyosin ring instability, making these cells a relevant model for studying mitotic dysfunction. Given the HPV-18-positive background of HeLa cells, the DYRK4 knockout model provides insights into how oncogenic viral proteins may intersect with cell division machinery, offering a system to explore potential therapeutic vulnerabilities in cervical cancer and other malignancies characterized by aberrant cytokinesis.
Researchers can employ DYRK4 Knockout HeLa Polyclonal Cells in a variety of experimental approaches, including high-content imaging screens for cytokinesis inhibitors, immunofluorescence microscopy of midbody markers such as MKLP1 and Aurora B, and multinucleation assays using DAPI staining. This knockout model also supports time-lapse live-cell imaging to track mitotic progression, western blotting to assess septin phosphorylation status, and flow cytometry for DNA content analysis to quantify polyploid populations. Rescue experiments with wild-type DYRK4 can validate specific phenotypic linkages. For further information or to request a quote, please contact Ascent Research.
