CCDC138 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the CCDC138 gene in the human HT29 colorectal adenocarcinoma cell line. This heterogeneous knockout model avoids clonal selection artifacts and provides a robust platform for investigating loss-of-function effects on mitotic regulation and colorectal cancer biology. The polyclonal nature ensures a broad representation of genetic alterations, enabling the study of diverse cellular responses to CCDC138 ablation.
The HT29 cell line, originally derived from the colorectal adenocarcinoma of a 44-year-old Caucasian female, serves as a well-established model for intestinal epithelium and colorectal tumorigenesis. These cells exhibit epithelial morphology and retain key signaling pathways implicated in cancer progression, making them a relevant system for mechanistic and pharmacological studies.
CCDC138 encodes a coiled-coil domain protein that functions as a critical regulator of mitotic progression. It directly interacts with PLK1 and alpha-tubulin, influencing PLK1 stability and kinase activity. This modulation affects downstream effectors CDK1, Cyclin B1, and Aurora A, which are crucial for mitotic entry, centrosome duplication, and spindle assembly. CCDC138 disruption thus impairs PLK1-mediated signaling, leading to aberrant spindle formation and delayed mitosis. The protein operates within the PLK1?CCyclin B1?CCDK1 axis and the Aurora A?CCDC25C pathway.
In HT29 colorectal cancer cells, CCDC138 loss compromises PLK1 function, causing genomic instability and reduced proliferation. This phenotype highlights colorectal cancer’s dependence on mitotic regulators and positions CCDC138 as a vulnerability. The polyclonal knockout population captures gene disruption heterogeneity, providing a relevant model for studying mitotic checkpoint control and PLK1 dysregulation in cancer.
This knockout model is suitable for Western blotting of PLK1 and phospho-histone H3, immunofluorescence of mitotic spindles, flow cytometry cell cycle analysis, colony formation and proliferation assays, and time-lapse microscopy. It supports PLK1 inhibitor research, functional genomics, cell cycle synchronization, and centrosome studies. For additional details, please contact Ascent Research.
