CCDC120 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human colorectal adenocarcinoma HT29 cell line, providing a genetically heterogeneous pool of cells with disrupted CCDC120 gene function through CRISPR/Cas9-mediated gene editing. This polyclonal format captures a range of loss-of-function events, offering a robust tool to study CCDC120 biology without the selection bias of a single clone.
HT29 cells are a widely used human colorectal adenocarcinoma epithelial cell line that produces mucin, representing a model of intestinal epithelium and colorectal cancer. These cells exhibit differentiated characteristics and are extensively employed to investigate signal transduction, cell adhesion, proliferation, and tumorigenesis mechanisms. Their capacity to form primary cilia makes them particularly relevant for studying ciliary protein functions in cancer.
CCDC120 encodes a centriolar protein essential for primary cilium formation, localizing to subdistal appendages of the mother centriole. It directly interacts with ODF2, CEP170, CEP89, and ninein to coordinate ciliogenesis. Loss of CCDC120 disrupts primary cilia, thereby impairing Hedgehog and Wnt signaling pathways. Downstream effects include reduced activation of GLI transcription factors and ??-catenin/TCF-dependent transcription, as well as altered AKT signaling. Key pathway components affected include SMO, GLI1, PTCH1, ??-catenin, and TCF7L2, with upstream regulation by RFX transcription factors and Foxj1.
Within HT29 colorectal cancer cells, CCDC120 knockout impairs ciliogenesis and associated signaling, providing a unique system to examine the interplay between primary cilia and colorectal tumor biology. Defective Hedgehog and Wnt signaling can influence cell proliferation, differentiation, and migration, mirroring aspects of colorectal cancer progression and ciliopathies. This model enables detailed dissection of how ciliary proteins modulate oncogenic pathways and maintain the transformed phenotype.
This knockout cell pool is ideally suited for a broad range of functional studies. Researchers can employ immunofluorescence microscopy with antibodies against acetylated tubulin or ARL13B to visualize cilia, complemented by western blotting for ciliary markers such as IFT88. Transcriptional analyses via RT-qPCR for Hedgehog target genes like GLI1 and PTCH1 can assess pathway activity. Functional assays including wound healing, cell proliferation, and colony formation enable interrogation of oncogenic properties. These tools support mechanistic studies of ciliary proteins in colorectal cancer biology. For further technical information, please contact Ascent Research.
