The DYNC2H1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HeLa cells with targeted disruption of the DYNC2H1 gene. This heterogeneous pool provides a loss-of-function model for studying cytoplasmic dynein-2 in retrograde intraflagellar transport (IFT) and ciliary signaling, without clonal isolation. The polyclonal format offers a versatile tool for functional genomics in cilia biology and disease modeling.
HeLa cells, derived from a human cervical adenocarcinoma, are HPV18-positive and immortalized, serving as a classical model for cervical cancer and epithelial cell biology. Under serum deprivation, HeLa cells can form primary cilia, enabling studies of ciliogenesis and IFT in a cancer context. This dual utility makes HeLa an ideal host for investigating ciliary protein function and oncogenic signaling interplay.
DYNC2H1 encodes the heavy chain of cytoplasmic dynein-2, the motor driving retrograde IFT in primary cilia. This dynein complex transports cargo from the ciliary tip to the cell body, a process crucial for ciliary assembly and hedgehog signaling. DYNC2H1 interacts with adaptors DYNC2LI1, WDR34, and WDR60 and coordinates with IFT-A and IFT-B complexes. Its activity is regulated by RFX transcription factors and cell cycle cues, and it functions downstream of kinesin-2 anterograde transport. DYNC2H1 disruption impairs retrograde IFT, leading to defective ciliogenesis, accumulation of ciliary membrane proteins, and reduced hedgehog pathway activity, as evidenced by diminished GLI transcription factor output. This mechanistic linkage underlies its association with ciliopathies such as short-rib polydactyly syndromes and Jeune syndrome.
In HeLa cells, DYNC2H1 knockout creates a powerful model to dissect ciliary dysfunction within an oncogenic background. The HPV18 E6/E7-expressing environment allows exploration of how impaired retrograde IFT and hedgehog signaling may influence cancer cell behaviors like proliferation and migration. This polyclonal knockout population enables investigation of ciliopathy mechanisms and the crosstalk between primary cilia and tumorigenic pathways, offering a unique platform to study the dual roles of DYNC2H1 in development and disease.
These polyclonal knockout cells support a range of applications including ciliopathy modeling, intraflagellar transport analysis, and hedgehog signaling studies. Researchers can employ immunofluorescence for ciliary markers (ARL13B, acetylated tubulin), western blotting for DYNC2H1, and RT-qPCR for GLI1 to characterize the knockout phenotype. Further functional assays like cell cycle and migration analyses can assess broader cellular impacts. These cells are a ready-to-use resource for cilia and cancer research. For ordering and inquiries, please contact Ascent Research.
