CCDC88C Knockout HEK293T Polyclonal Cells are a polyclonal knockout cell population derived from HEK293T cells, generated through CRISPR/Cas9-mediated disruption of the CCDC88C gene. This pooled knockout model provides a heterogeneous collection of edited cells, each carrying distinct mutations at the target locus, reflecting the spectrum of genomic modifications induced by non-homologous end joining repair. As a polyclonal knockout product, it is suited for population-level assays where bulk loss-of-function effects are interrogated, avoiding potential clonal artifacts while maintaining the genetic diversity inherent in the edited pool.
The parental HEK293T cell line is a human embryonic kidney epithelial derivative that constitutively expresses the SV40 large T antigen, a feature that enables high-copy episomal replication of plasmids containing the SV40 origin of replication. This property, together with its exceptional transfectability, makes HEK293T a preferred host for recombinant protein expression, lentivirus production, and functional genomics experiments. The cell line retains active Wnt, MAP kinase, and Rho GTPase pathways, offering a versatile background for genetic perturbation studies.
CCDC88C, also designated Daple, encodes a multidomain scaffolding protein that occupies a central node in non-canonical Wnt/planar cell polarity (PCP) signaling. Mechanistically, it forms a complex with Dishevelled (DVL1/2/3) and the core PCP components VANGL1/2 and PRICKLE1, and subsequently couples these proximal signaling events to the activation of RhoA, Rac1, and JNK. Through this coupling, CCDC88C translates extracellular WNT5A and WNT11 signals, received by Frizzled receptors, into dynamic rearrangements of the actin cytoskeleton. This pathway is crucial for directed cell migration, establishment of epithelial polarity, and morphogenetic processes such as neural tube closure.
Within the HEK293T context, the CCDC88C knockout polyclonal cells permit dissection of Rho GTPase-driven signaling events in a model system that is both readily transfectable and well-characterized for Wnt pathway studies. Because HEK293T cells do not exhibit complex epithelial polarization programs, the phenotype attributable to Daple loss can be attributed more directly to cytoskeletal and migration defects rather than to developmental polarity cues. Researchers can rescue the knockout phenotype by expressing wild-type or mutant CCDC88C constructs, enabling structure-function analyses. Moreover, the cell line??s robust growth and compatibility with high-content imaging platforms facilitate scalable screening campaigns.
Key experimental applications include quantitative assessment of cell migration using wound healing and transwell invasion assays, visualization of F-actin architecture via immunofluorescence, and biochemical determination of RhoA and Rac1 activation status through pull-down approaches. Co-immunoprecipitation experiments can confirm the disruption of the CCDC88C-Dishevelled interaction, while phospho-specific western blotting for JNK provides a readout of pathway activity. These applications are central to investigations of cancer cell dissemination, neural tube defect etiology, and Wnt/PCP signal transduction. For further details or ordering inquiries, please contact Ascent Research.
