The DNAH5 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric carcinoma cell line HGC-27. This model introduces targeted disruption of the DNAH5 gene, which encodes the dynein axonemal heavy chain 5 protein required for ciliary motility. The polyclonal format provides a heterogeneous edited pool, avoiding clonal selection bottlenecks and enabling robust loss-of-function studies. This product is ideal for dissecting DNAH5-dependent processes in ciliary biology and gastric cancer research.
The HGC-27 cell line originates from a lymph node metastasis of a gastric adenocarcinoma, serving as a widely used model for metastatic gastric cancer. These epithelial cells can form motile cilia, and DNAH5 has been shown to undergo epigenetic silencing via promoter methylation in gastric tumors. Thus, knocking out DNAH5 in HGC-27 cells creates a physiologically relevant system to study how ciliary dysfunction contributes to cancer cell behavior and signaling, mirroring clinical observations of DNAH5 suppression in gastric malignancies.
DNAH5 functions as an ATP-driven motor protein within outer dynein arms of ciliary axonemes, interacting with DNAI1, DNAI2, DNAL1, and acetylated tubulin to generate ciliary beating. Its expression is governed by FOXJ1 and RFX transcription factors downstream of NOTCH signaling. Knockout of DNAH5 abolishes ciliary motility, impairing cilia-generated fluid flow and dampening downstream Hedgehog and PDGF signaling pathways. This disruption highlights the gene’s central role in mechanotransduction and mucociliary clearance.
In the gastric cancer context, DNAH5 loss in HGC-27 cells recapitulates the common epigenetic silencing observed in tumors, enabling functional studies on how ciliary motor defects influence migration and signaling. This polyclonal knockout model allows assessment of heterogeneous responses to ciliary ablation, more closely reflecting in vivo tumor heterogeneity. It provides a platform to interrogate the role of cilia in metastasis-related processes such as wound healing, directional migration, and growth factor signaling.
Applications include modeling primary ciliary dyskinesia, analyzing DNAH5 methylation patterns, and performing mucociliary clearance assays. Researchers can evaluate ciliary structure via immunofluorescence for acetylated tubulin, measure ciliary beat frequency by video microscopy, and quantify gene expression with qPCR. Scratch wound assays assess migration, while methylation-specific PCR probes epigenetic status. This knockout cell population is a versatile tool for elucidating cilia-dependent mechanisms in cancer and ciliopathies. For further details, contact Ascent Research.
