The DCLK1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T cell line, featuring targeted disruption of the doublecortin-like kinase 1 (DCLK1) gene. This polyclonal pool provides a robust loss-of-function model for investigating DCLK1-dependent biological mechanisms without the selective pressures associated with single-cell cloning. The knockout is generated through CRISPR/Cas9-mediated gene disruption, enabling efficient ablation of DCLK1 expression across the cell population. This product is suitable for a range of functional studies exploring the kinase??s role in cellular processes, particularly those relevant to cancer stem cell biology and microtubule regulation.
Host cell background: HEK293T cells are a human embryonic kidney line stably expressing the SV40 large T antigen, which permits episomal replication of plasmids containing the SV40 origin of replication. This feature confers high transfection efficiency and robust protein expression capacity, making HEK293T an industry-standard host for viral production, recombinant protein expression, and general cell biology studies. The well-characterized nature of HEK293T cells provides a consistent and reproducible platform for gene-editing applications. Their rapid proliferation and ease of culture further enhance their suitability for creating knockout models that require scalable cell populations for diverse assay formats.
DCLK1 encodes a microtubule-associated serine/threonine kinase that regulates key signaling nodes implicated in neuronal migration and oncogenic stemness. At the molecular level, DCLK1 is activated by upstream Wnt ligands and TCF/LEF transcription factors, as well as the Notch intracellular domain, thereby integrating canonical Wnt/??-catenin and Notch pathway inputs. Its downstream effectors include the pluripotency transcription factors SOX2 and NANOG, along with epithelial-mesenchymal transition (EMT) markers such as SNAI1 and TWIST1, all of which are critical for stem cell maintenance and tumor progression. DCLK1 directly interacts with microtubules and tubulin, influencing cytoskeletal dynamics, and participates in multiprotein complexes with other kinases. Disruption of DCLK1 abrogates its kinase activity and destabilizes these signaling networks, leading to impaired regulation of gene programs that control cell migration, self-renewal, and differentiation. Representative pathway components encompass Wnt ligands, Frizzled receptors, ??-catenin, TCF/LEF, Notch receptors, ??-secretase, and RBP-J??.
In the HEK293T cellular background, DCLK1 knockout offers a defined model to dissect DCLK1??s contribution to Wnt and Notch signal transduction without the confounding endogenous mutations often found in cancer cell lines. The embryonic kidney origin and high transfectability of this line allow for precise reconstitution experiments and reporter-based readouts, facilitating structure-function analyses of DCLK1 domains. Researchers can use this model to assess how loss of DCLK1 alters microtubule organization, subcellular localization of pathway effectors, and transcriptional outputs. Additionally, the absence of DCLK1 may sensitize cells to pharmacological inhibitors, enabling drug target validation studies. The polyclonal nature of the population preserves genetic heterogeneity, providing a realistic backdrop for evaluating phenotypic variability upon target-gene perturbation.
Applications for this product span a wide range of biomedical research areas, including cancer stem cell biology, neurodevelopmental signaling, and microtubule dynamics. Typical experimental workflows include western blotting to confirm DCLK1 knockdown, immunofluorescence staining to visualize changes in microtubule architecture, and migration/invasion assays to quantify metastatic potential. RT-qPCR for stemness markers (SOX2, NANOG) and EMT regulators (SNAI1, TWIST1) can reveal transcriptional consequences of DCLK1 loss, while Wnt/??-catenin and Notch signaling reporter assays illuminate pathway activity. These cells are also suitable for co-culture studies and high-throughput screens to identify synthetic lethal interactions or novel DCLK1 modulators. For further details on lot-specific performance or customization options, please contact Ascent Research.
