The CCDC93 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HeLa cells with targeted disruption of the CCDC93 gene. This pool offers a heterogeneous loss-of-function model that eliminates functional CCDC93 protein, enabling study of CCDC93-dependent processes without clonal bias. The polyclonal format is ideal for experiments requiring a robust null background while maintaining genetic diversity.
The host HeLa line is an immortalized human cervical adenocarcinoma epithelial cell line positive for HPV-18. Commonly used to model epithelial cell biology and oncogenesis, HeLa cells possess well-defined endocytic and recycling pathways, making them an optimal platform for investigating retromer-independent receptor trafficking.
CCDC93 is an essential subunit of the CCC complex, which localizes to early endosomes and coordinates retromer-independent recycling of receptors such as LDLR. Together with CCDC22 and COMMD1-10, the CCC complex engages the retriever module (VPS35L, VPS26C, VPS29) and SNX17 to sort internalized cargo away from lysosomal degradation. Regulation by endosomal phosphatidylinositol-3-phosphate and COMMD proteins controls CCDC93 activity, which directly governs LDLR surface presentation, cholesterol internalization, and nutrient receptor homeostasis. Disruption of CCDC93 impairs receptor recycling and disrupts cholesterol balance, underscoring its centrality in receptor-mediated endocytosis.
In HeLa cells, CCDC93 ablation creates a valuable model to explore retromer-independent endosomal sorting within a malignant epithelial background. HeLa cells exhibit active LDLR trafficking and cholesterol metabolism, allowing researchers to dissect how the CCC complex sustains receptor abundance and metabolic fitness in cancer. This knockout system can link endosomal trafficking deficiencies to mechanisms underlying familial hypercholesterolemia and cardiovascular pathology, and may reveal cancer-specific dependencies on nutrient receptor recycling for growth.
These polyclonal knockout cells support a range of assays, including Western blotting and flow cytometry to quantify LDLR protein levels, LDL uptake assays to measure receptor function, and immunofluorescence microscopy to visualize endosomal marker distribution. Co-immunoprecipitation validates CCC/retriever/SNX17 interactions, while cholesterol efflux assays and RNA-seq provide broader metabolic and transcriptomic insights. Applications span cholesterol metabolism studies, high-throughput LDLR modulator screening, and exploration of endosomal trafficking in cancer biology. For additional information, please contact Ascent Research.
