The DNAH5 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell carcinoma cell line. This product features targeted disruption of the DNAH5 gene, which encodes the dynein axonemal heavy chain 5, an essential component of the outer dynein arm complex required for ciliary motility. The polyclonal nature ensures a heterogeneous mixture of edited alleles, providing a robust loss-of-function model without clonal isolation, and is suitable for a broad range of functional studies where complete gene knockout is not a prerequisite. The cells are offered as a ready-to-use tool for investigating cilia biology, signaling, and disease mechanisms in a cancer-relevant context.
The 786-O parental cell line is a well-established model of clear cell renal cell carcinoma (ccRCC) originating from a primary tumor. These cells harbor a characteristic von Hippel-Lindau (VHL) gene mutation, which leads to constitutive activation of hypoxia-inducible factor (HIF) signaling and mimics key aspects of ccRCC pathogenesis. As a renal epithelial tumor model, 786-O cells grow adherently and are widely employed to study VHL-dependent pathways, tumor metabolism, and cellular responses to hypoxia. The introduction of DNAH5 knockout into this background provides a unique platform to examine the intersection of ciliary dysfunction and oncogenic signaling in kidney cancer.
DNAH5 encodes the heavy chain subunit of the outer dynein arm, an ATPase-powered motor protein that drives rhythmic beating of motile cilia and flagella. In the ciliary axoneme, DNAH5 forms complexes with other outer dynein arm subunits including DNAI1, DNAI2, and DNAL1. Upstream transcription factors FOXJ1 and members of the RFX family directly regulate DNAH5 expression, coordinating ciliogenesis programs. Downstream, functional outer dynein arms enable ciliary motility, which is critical for mucociliary clearance, fluid flow sensing, and transduction of the Hedgehog (Hh) signaling pathway. Specifically, ciliary movement facilitates proteolytic processing of GLI transcription factors, thereby modulating Hh target gene expression. Disruption of DNAH5 impairs ciliary beat frequency and dampens flow-dependent signaling cascades, making these cells an ideal model to dissect motile cilia-dependent signaling events.
The combination of VHL-mutant renal carcinoma background and DNAH5 knockout creates a powerful system to investigate roles of primary and motile cilia in kidney cancer biology. While 786-O cells typically possess primary cilia, the loss of DNAH5 may perturb ciliary structure or function, potentially affecting Hedgehog signaling dynamics that are already altered by VHL loss. This model enables researchers to assess how ciliary motility defects influence tumor cell behavior, including migration, invasion, and response to microenvironmental cues. Moreover, it provides an opportunity to study the poorly understood contributions of ciliary genes to renal cell carcinoma progression and metastasis.
Typical research applications include studying ciliogenesis and ciliary protein trafficking, modeling primary ciliary dyskinesia and Kartagener’s syndrome phenotypes, and investigating cilia-dependent Hedgehog pathway activation using GLI reporter assays. The cells are also suitable for drug screening to identify small molecules that rescue ciliary motility or modulate downstream signaling. Representative experimental techniques include immunofluorescence staining for ciliary markers (e.g., acetylated ??-tubulin, ARL13B), high-speed video microscopy for ciliary beat frequency measurement, western blotting for DNAH5 protein expression, RT-qPCR analysis of ciliogenesis regulators like FOXJ1 and RFX2, and cell migration assays to evaluate functional consequences of ciliary loss. For further information, please contact Ascent Research.
