The DPCD Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a mixed population with targeted disruption of the DPCD gene, enabling loss-of-function studies in a genetically diverse background. Unlike clonal cell lines, polyclonal populations retain heterogeneity that can better represent physiological variation. The knockout cells are designed for researchers investigating ciliary biology, epithelial signaling, and related disease models. As polyclonal knockout cells, they are suitable for pooled functional assays without the need for single-cell cloning.
The parental HT29 cell line originates from a human colorectal adenocarcinoma and serves as a well-established model of intestinal epithelial biology. These cells exhibit characteristic epithelial morphology, express markers such as villin and mucin, and can undergo differentiation into enterocyte-like cells with brush border features. HT29 cells are extensively utilized to study intestinal barrier function, cell polarity, and differentiation pathways. Importantly, HT29 cells possess primary cilia, making them a relevant system for probing ciliary structure and function in an epithelial context. Their tumorigenic origin also permits investigation of oncogenic signaling interactions with ciliary pathways.
DPCD (Deleted in Primary Ciliary Dyskinesia) encodes a critical component of the nexin-dynein regulatory complex (N-DRC), which is essential for ciliary motility and axonemal integrity. DPCD protein interacts with other N-DRC subunits such as DRC2, DRC3, and DRC4 to modulate dynein arm activity and control ciliary beat frequency. Its expression is regulated by transcription factors RFX and FOXJ1, master regulators of ciliogenesis. Downstream, DPCD influences mucociliary clearance and the Hedgehog signaling cascade through effects on GLI transcription factors. Disruption of DPCD thus compromises ciliary movement and signal transduction, linking primary ciliary dyskinesia to defects in organ laterality and chronic respiratory infections.
In the HT29 background, DPCD knockout generates a physiologically relevant model to dissect the interplay between ciliary function and intestinal epithelial homeostasis. Loss of DPCD is expected to impair ciliary ultrastructure and dampen cilia-dependent Hedgehog signaling, potentially altering differentiation programs and barrier integrity. Given HT29’s capacity for mucin production, this model enables examination of how ciliary defects affect glycosylation and mucus layer properties. Furthermore, the disruption of the N-DRC may uncouple Wnt/planar cell polarity cues, providing insight into tissue patterning mechanisms. This system thus bridges ciliopathy research with colorectal epithelial biology, offering a platform to study diseases like primary ciliary dyskinesia in a tractable cell-based format.
Researchers can employ these polyclonal knockout cells in a wide range of assays, including immunofluorescence staining for ciliary markers (e.g., acetylated tubulin, DRC2), western blotting to confirm loss of N-DRC components, and RT-qPCR for ciliary gene expression. Functional evaluations such as ciliary beat frequency analysis using high-speed microscopy and mucociliary transport assays are feasible. Additionally, cell differentiation studies monitoring alkaline phosphatase activity and mucin secretion can reveal phenotypic consequences. Proliferation and drug screening assays enable testing of therapeutic candidates for ciliopathies. For detailed technical specifications and ordering information, please contact Ascent Research.
