The DNAAF2 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated by targeted disruption of the DNAAF2 gene in the SK-HEP-1 human cell line. This polyclonal pool comprises a heterogeneous mixture of cells carrying various loss-of-function mutations at the DNAAF2 locus, providing a robust model for studying gene function without the limitations of clonal selection. The use of polyclonal knockout populations mitigates clonal artifacts and allows for more representative analysis of gene disruption effects in a cellular context.
The SK-HEP-1 cell line is derived from ascitic fluid of a patient with liver adenocarcinoma and is widely utilized as a model for hepatic sinusoidal endothelium due to its endothelial characteristics. These adherent epithelial-like cells exhibit expression of endothelial markers and are capable of forming primary cilia, making them a valuable platform for investigating ciliary biology in a hepatic endothelial milieu. Their dual hepatic and endothelial features enable studies linking liver pathology with ciliopathy-associated phenotypes.
DNAAF2 functions as a cytoplasmic co-chaperone that cooperates with HSP70 and HSP90 to fold axonemal dynein heavy chains, such as DNAH5 and DNAH11, facilitating their preassembly into dynein arm complexes critical for motile ciliary function. The expression of DNAAF2 is transcriptionally regulated by FOXJ1, RFX2, and RFX3, master regulators of ciliogenesis. DNAAF2 interacts with other dynein assembly factors including DNAAF1 and DNAAF3, forming a chaperone network essential for the cytoplasmic preassembly of outer and inner dynein arms. Its disruption impairs intraflagellar transport and leads to defects in ciliary motility.
In the SK-HEP-1 background, DNAAF2 knockout provides a physiologically relevant system to dissect ciliogenesis and dynein arm assembly in cells that model the hepatic sinusoidal endothelium. Given the emerging link between ciliary dysfunction and hepatic diseases, this model allows researchers to explore whether motile cilia defects contribute to liver pathophysiology. It offers a valuable tool for primary ciliary dyskinesia research within an endothelial context, bridging the gap between respiratory, reproductive, and hepatic manifestations of ciliopathies.
This knockout product supports diverse experimental applications, including biochemical analysis of dynein arm components via Western blotting and co-immunoprecipitation, visualization of ciliary markers (Arl13b, acetylated-tubulin) by immunofluorescence, transcriptional profiling of ciliogenesis genes through RT-qPCR, and ultrastructural examination of axonemal architecture using transmission electron microscopy. It is suitable for drug screening efforts targeting ciliopathy-related pathways and for functional studies of DNAAF2 in motile cilia assembly. For further information or technical support, please contact Ascent Research.
