DNAH5 Knockout Huh-7 Polyclonal Cells is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNAH5 gene in the human Huh-7 hepatocellular carcinoma cell line. This versatile tool enables loss-of-function studies of DNAH5, which encodes an axonemal dynein heavy chain essential for ciliary beat generation and mucociliary clearance. The polyclonal nature retains genetic heterogeneity while providing robust knockout efficiency for population-level analyses. No specific mutation or editing mechanism is claimed; the product is designed for researchers seeking to disrupt DNAH5 expression and study its downstream effects.
Huh-7 cells are a well-differentiated, epithelial hepatocellular carcinoma line derived from a human male. They serve as a widely used hepatocyte model for drug metabolism, HCV replication, and hepatic physiology. Their epithelial morphology and robust growth characteristics make them suitable for transfection, imaging, and biochemical assays. In the context of this knockout, Huh-7 cells provide a consistent genetic background to evaluate DNAH5 function and facilitate assay development.
DNAH5 encodes the heavy chain of the outer dynein arm, a component of the axonemal dynein motor complex that powers ciliary movement. It functions within the ciliary axoneme, interacting with structural partners such as DNAI1, DNAH11, and NME8, as well as radial spoke proteins and the outer dynein arm docking complex. Upstream, transcription factors FOXJ1, RFX, and Multicilin regulate DNAH5 expression, while its activity directly drives mucociliary clearance and impacts Hedgehog signaling indirectly through ciliary function. Knockout of DNAH5 leads to immotile cilia, modeling the pathogenesis of primary ciliary dyskinesia (PCD) and Kartagener syndrome.
In Huh-7 cells, the DNAH5 knockout provides a unique platform to dissect ciliopathy mechanisms in an epithelial context. Although Huh-7 cells are not motile-ciliated, they retain primary cilia and key signaling pathways, enabling investigation of DNAH5 protein interactions, CRISPR editing validation, and off-target assessments. This model is particularly useful as a negative control in ciliary motility studies and for examining the role of dynein arm components in cellular processes beyond ciliary beating, such as intracellular trafficking or signaling.
Researchers can employ this knockout for ciliopathy disease modeling, validating downstream effects through immunofluorescence to assess ciliary structure, Western blotting for protein expression changes, RT-qPCR for transcript analysis, and Sanger sequencing or targeted deep sequencing to confirm gene disruption. The polyclonal format supports population-based functional screens and drug testing. For additional information or to discuss customized options, please contact Ascent Research.
