CCDC186 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HeLa cells, providing a loss-of-function model for studying CCDC186. The polyclonal format yields a pool of cells with targeted gene disruption, suitable for bulk functional assays and pooled screening applications.
HeLa is a human cervical adenocarcinoma epithelial cell line with integrated HPV18 sequences, extensively used in cancer biology for investigating tumorigenesis and cell cycle dysregulation. Under serum starvation, HeLa cells assemble primary cilia, enabling studies of ciliary trafficking and Hedgehog signaling in a cancer-relevant setting.
The CCDC186 protein is an essential component of retrograde intraflagellar transport (IFT), facilitating the return of signaling proteins from the ciliary tip. It interacts with IFT88, IFT140, and BBSome subunits within IFT-A/B complexes. Its expression is controlled by RFX transcription factors and FOXJ1. Loss of CCDC186 disrupts ciliary localization of Smoothened (SMO) and GPCRs such as SSTR3 and HTR6, leading to impaired activation of GLI transcription factors and altered expression of Hedgehog targets like GLI1 and PTCH1.
In HeLa cells, CCDC186 knockout perturbs cilia-dependent signaling, offering a platform to investigate Hedgehog pathway dysregulation in cancer. Given the HPV18-driven oncogenic background, this model can be used to explore how viral oncoproteins modulate ciliary function and how ciliary defects influence tumor cell proliferation and survival. Although CCDC186 mutations are associated with ciliopathies such as neurodevelopmental disorder with obesity and retinal dystrophy, this knockout model in a cancer cell line provides insights into the broader role of ciliary trafficking in cellular homeostasis and disease.
Typical applications include ciliogenesis assays under serum starvation, immunofluorescence detection of ciliary markers ARL13B and acetylated tubulin, SMO translocation assays, and western blot analysis of GLI3 processing to assess full-length and repressor forms. Transcriptional profiling of Hedgehog target genes GLI1 and PTCH1 by RT-qPCR, along with co-immunoprecipitation of IFT components and flow cytometry for ciliated cell quantification, provide robust readouts for pathway activity. These polyclonal knockout cells are also valuable for screening chemical or genetic modulators of ciliogenesis and GPCR ciliary signaling. For further information, please contact Ascent Research.
