The CCDC127 Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal HeLa cell population with targeted disruption of the CCDC127 gene, which encodes a centriolar satellite protein critical for ciliogenesis and microtubule organization. This knockout model provides a genetically heterogeneous pool where loss of CCDC127 function can be studied across a range of alleles, facilitating analysis of gene function in a human cervical adenocarcinoma background.
The HeLa host cell line is an immortalized human cervical adenocarcinoma model extensively used in cancer biology, cell signaling, and drug discovery. Under serum-free conditions, HeLa cells can form primary cilia, making them a suitable platform for investigating ciliogenesis and the hedgehog signaling cascade. Their well-characterized genome and robust growth properties further support reproducible functional assays.
At the molecular level, CCDC127 localizes to centriolar satellites where it forms complexes with scaffolding proteins PCM1, CEP290, and CEP131 to orchestrate microtubule-dependent trafficking of ciliary components. Its function is tightly regulated by cell cycle-dependent kinases, including PLK1 and Aurora A, linking ciliogenesis to proliferative signals. Downstream, CCDC127 activity is essential for proper hedgehog pathway transduction; it facilitates the localization and activation of SMO, the GLI transcription factors (GLI1, GLI2, GLI3), and intraflagellar transport machinery such as IFT88. The pathway is further modulated by the negative regulator SUFU, which is also affected by CCDC127 loss. Consequently, knockout of CCDC127 impairs primary cilium assembly and dampens hedgehog target gene expression.
In the HeLa cervical cancer context, CCDC127 deficiency disrupts the formation of primary cilia and attenuates hedgehog signaling, providing a defined cellular model to dissect the interplay between centrosomal proteins, ciliogenesis, and tumor biology. This system is particularly relevant for studying ciliopathies and for examining how ciliary defects influence cancer cell phenotypes, including altered migration, proliferation, and cell cycle progression. Additionally, it enables screening of modulators that may restore cilia function or hedgehog activity.
Experimentally, these polyclonal knockout cells support a variety of applications: immunofluorescence staining for ARL13B and acetylated tubulin to visualize cilia and quantify cilium length, western blotting to assess levels of hedgehog pathway proteins (e.g., GLI1, SMO), and RT-qPCR analysis of downstream transcriptional targets. Functional assays such as wound-healing migration tests and cell cycle profiling further reveal the consequences of CCDC127 loss. Altogether, the model is ideal for detailed investigations in ciliogenesis, centrosome biology, and cancer cell signaling. For further information or technical assistance, please contact Ascent Research.
