The DNAH5 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from the SK-HEP-1 human hepatic sinusoidal endothelial cell line, carrying a targeted disruption of the DNAH5 gene. DNAH5 encodes an axonemal dynein heavy chain essential for the assembly of outer dynein arms in motile cilia, and its loss of function serves as a genetically defined model for primary ciliary dyskinesia and related ciliopathies. This product provides a mixed knockout pool rather than a monoclonal line, enabling the study of gene disruption effects across a spectrum of allelic variants without fixation on a single clone.
The host cell model, SK-HEP-1, is an established adherent line originally derived from a human hepatic adenocarcinoma and extensively characterized as a surrogate for liver sinusoidal endothelial cells. These cells natively fulfill roles in blood filtration, metabolic exchange, and sinusoidal lining, and they have been adapted for diverse biomedical research applications. Although SK-HEP-1 cells are not classical ciliated epithelia, they can express components of the ciliary machinery under appropriate conditions, making them a tractable system for investigating motile cilia gene function in a readily manipulable human cell background.
At the molecular level, DNAH5 functions as a core component of the outer dynein arm, generating ATP-dependent force for ciliary beat. Its transcription is regulated by key ciliogenesis factors including FOXJ1, RFX2, RFX3, and MCIDAS, while the assembled protein complex interacts with other outer dynein arm constituents such as DNAH11, DNAI1, DNAI2, and DNALI1. Disruption of DNAH5 abolishes outer dynein arm formation, leading to impaired ciliary motion, reduced beat frequency, and downstream failures in mucociliary clearance, fluid flow generation, and establishment of left?Cright body asymmetry??hallmarks of Kartagener syndrome and primary ciliary dyskinesia.
In the context of SK-HEP-1 knockout cells, DNAH5 ablation creates a loss-of-function model that allows researchers to dissect the structural and functional consequences of outer dynein arm deficiency without the complexity of primary ciliated cell culture. This cellular background facilitates biochemical analyses of dynein complex assembly and stability, as well as live-cell imaging of residual ciliary dynamics. The knockout pool can be used to screen for pharmacological rescue agents or to introduce wild-type and mutant DNAH5 constructs for structure?Cfunction studies, providing a flexible platform complementary to animal models and patient-derived samples.
Typical applications include high-speed video microscopy to measure ciliary beat frequency, immunofluorescence staining for localization of ciliary markers (e.g., acetylated ??-tubulin, DNAI1), western blotting for outer dynein arm protein levels, RT-qPCR profiling of ciliogenesis transcription factors, and mucociliary clearance assays. This product is particularly suited for functional genomics screening, ciliopathy drug discovery, and investigation of cilia-dependent signaling pathways. For detailed inquiries or to discuss custom applications, please contact Ascent Research.
