The DNAH5 knockout AGS polyclonal cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the AGS human gastric adenocarcinoma epithelial cell line, featuring targeted disruption of the DNAH5 gene. This loss-of-function model provides researchers with a powerful tool to investigate the role of DNAH5 in ciliary motility and gastric epithelial biology. The polyclonal nature of the knockout pool ensures a heterogeneous genetic background, enabling robust assessment of gene function in a mixed population and reducing clonal bias in downstream experiments.
The AGS cell line was established from a gastric adenocarcinoma and is widely employed as a model for gastric epithelial signaling, disease pathogenesis, and host?Cmicrobe interactions. These epithelial cells retain key characteristics of gastric mucosal cells, including the ability to form primary cilia and, under certain conditions, motile cilia. Their stable growth properties and well-characterized molecular landscape make AGS cells an ideal host for studying ciliary gene function in a gastrointestinal context, particularly given the emerging role of ciliary dysfunction in cancer and chronic inflammation.
DNAH5 encodes dynein axonemal heavy chain 5, a core ATPase subunit of the outer dynein arm complex in motile cilia and flagella. Expression of DNAH5 is controlled by the transcription factors FOXJ1 and RFX2, which orchestrate ciliogenesis, while NOTCH signaling influences ciliated cell fate. Within the axoneme, DNAH5 forms functional interactions with DNAI1, DNAH11, and DNAL1, and its proper folding depends on the molecular chaperones HSP40 and HSP70. The ATPase activity of DNAH5 directly generates force for ciliary beating; thus, it is a key determinant of ciliary beat frequency, which drives mucociliary clearance and establishes left-right asymmetry during embryogenesis.
Disruption of DNAH5 in AGS cells mimics the molecular deficit observed in primary ciliary dyskinesia (PCD), a disorder characterized by chronic respiratory infections, sinusitis, bronchiectasis, and situs inversus. In the gastric epithelium, impaired ciliary function due to DNAH5 loss can diminish mucociliary clearance, potentially altering the interaction between gastric epithelial cells and pathogens such as Helicobacter pylori. This model therefore provides a unique platform to dissect how ciliary defects contribute to gastric epithelial dysfunction, chronic inflammation, and susceptibility to microbial colonization, bridging ciliopathy research with gastroenterology.
This knockout polyclonal population is suited for diverse applications. Ciliary structure and function can be examined by immunofluorescence for axonemal markers (e.g., acetylated ??-tubulin) and high-speed video microscopy to measure ciliary beat frequency. Western blotting for DNAH5, RT-qPCR for cilia-related genes, and RNA-seq transcriptomic profiling enable molecular characterization. Functional assays such as cell migration, invasion, and flow cytometry for surface markers further probe cilia-dependent phenotypes in gastric cancer. Drug screening for primary ciliary dyskinesia and host-pathogen interaction studies with H. pylori are additional uses. For further information, contact Ascent Research.
