The CCDC97 Knockout HT29 Polyclonal Cells provide a polyclonal CRISPR/Cas9-edited knockout cell population in the HT29 human colorectal adenocarcinoma cell line. This loss-of-function model relies on CRISPR/Cas9-mediated disruption of the CCDC97 gene, which encodes a coiled-coil domain-containing protein essential for centrosome function and ciliogenesis. The polyclonal format comprises a heterogeneous pool of edited cells, enabling population-level studies of gene disruption without clonal isolation. Researchers investigating microtubule organization, centrosome biology, and cell cycle regulation will find this product a robust tool for probing CCDC97-dependent mechanisms.
HT29 cells derive from a human colorectal adenocarcinoma and display epithelial morphology, characteristic of intestinal epithelium. They serve as a well-established model for colorectal cancer and intestinal epithelial biology, retaining the capacity to form polarized monolayers and differentiate under defined conditions. Their extensive characterization includes defined signaling pathways and genetic markers relevant to colorectal carcinogenesis. Introduction of CCDC97 loss into this background allows dissection of centrosomal defects within a clinically relevant cancer context, providing insights into how centrosome integrity impacts tumor cell behavior.
CCDC97 localizes to centriolar satellites and interacts with key components such as CSPP1 and PCM1, linking it to centriole duplication, microtubule anchoring, and cilia formation. It functions within a network involving the kinase PLK4, the centriole assembly factors STIL, SAS6, CEP135, CPAP, CEP152, and CEP63, as well as the cell cycle regulators CDK1 and Aurora A. Disruption of CCDC97 impairs these interactions, leading to defective centrosome maturation, aberrant mitotic spindle assembly, G2/M transition arrest, and compromised ciliogenesis. Consequently, this knockout model disrupts the coordination between centrosome duplication and cell cycle progression, with downstream effects on microtubule cytoskeleton organization.
In the HT29 colorectal adenocarcinoma context, loss of CCDC97 is particularly relevant for examining how centrosome dysfunction contributes to tumorigenesis. Centrosome abnormalities are frequent in colorectal cancer and can promote genomic instability, altered proliferation, and impaired differentiation. By abolishing CCDC97, this model may highlight cilia-dependent signaling pathways, such as those regulated by Wnt or Hedgehog, which are known to influence intestinal epithelial homeostasis and cancer. Thus, researchers can explore the roles of centrosome integrity and ciliary signaling in colorectal cancer progression, migration, and invasion.
This knockout cell population is suitable for a range of experimental applications. Researchers can perform immunofluorescence to visualize centrosome and cilia markers, western blotting to assess cell cycle proteins, and flow cytometry to analyze DNA content. Proliferation assays such as MTT or BrdU, migration/invasion assays, and RNA-seq for transcriptomic profiling enable functional characterization. Co-immunoprecipitation studies can probe CCDC97 interactors like CSPP1 and PCM1. The model also supports screening of centrosome-targeted therapeutic compounds. For further technical details or ordering assistance, please contact Ascent Research.
