CCDC6 Knockout HEK293T Polyclonal Cells are a heterogeneous population of HEK293T cells bearing CRISPR/Cas9-mediated disruption of the human CCDC6 gene. This polyclonal knockout product provides a robust loss-of-function model for investigating CCDC6-dependent biological processes, including DNA damage response, microtubule dynamics, and ciliogenesis. The use of polyclonal cells rather than clonal isolates more closely reflects population-level behavior and mitigates clonal artifacts, making it suitable for a variety of downstream functional assays.
HEK293T cells are a widely used human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, enabling high-level plasmid amplification and transient protein overexpression. Their exceptional transfectability, rapid growth, and well-characterized signaling networks make them an ideal host for generating knockout models. The epithelial origin and retention of key kidney cell features also support studies in cellular morphology and epithelial biology.
CCDC6 encodes a coiled-coil domain-containing protein that functions at the intersection of genomic stability and cytoskeletal organization. In response to DNA double-strand breaks, CCDC6 is phosphorylated by the upstream kinases ATM and ATR, which promotes its interaction with and activation of the tumor suppressor p53, culminating in cell cycle arrest or apoptosis. Independently of its genotoxic stress role, CCDC6 localizes to microtubules and is required for microtubule stabilization and proper primary cilium assembly. In thyroid and lung cancers, chromosomal rearrangements fuse CCDC6 with the RET tyrosine kinase, generating oncogenic RET/PTC1 fusions that drive aberrant RET signaling. Thus, CCDC6 acts as a scaffold integrating ATM/ATR?Cp53 signaling with cytoskeletal and ciliary functions.
The HEK293T background is particularly advantageous for CCDC6 functional studies because these cells are readily transfected with DNA damage sensors, reporters, or wild-type CCDC6 constructs for rescue experiments. Their flat morphology and robust attachment to substrates facilitate high-resolution immunofluorescence imaging of microtubule networks, cilia formation, and ??H2AX foci. Moreover, HEK293T cells express a functional DNA damage response machinery, including ATM and ATR, allowing direct investigation of genotoxic stress pathways without the confounding mutations often found in cancer lines. Combined with the polyclonal knockout, researchers can assess population-level heterogeneity in DNA repair, apoptosis, and cell cycle regulation.
This CCDC6 knockout model is ideally suited for investigating DNA damage response kinetics and repair pathway choice using assays such as ??H2AX immunofluorescence, comet assay, and Western blotting for phosphorylated ATM/ATR substrates and p53. In cancer drug discovery, the polyclonal population enables screening of small-molecule inhibitors targeting the RET/PTC1 oncoprotein or modulators of p53-dependent apoptosis, with readouts including flow cytometry for DNA content and apoptosis markers. For ciliogenesis research, immunostaining for acetylated tubulin or ARL13B can visualize primary cilia defects. Additionally, RT-qPCR and Western blotting can quantify downstream gene expression changes after genotoxic challenge. For further details or custom orders, please contact Ascent Research.
