The DNAAF2 Knockout AGS Polyclonal Cells product provides a CRISPR/Cas9-edited heterogeneous population of AGS gastric epithelial cells carrying targeted gene disruption at the DNAAF2 locus. This polyclonal knockout cell pool enables loss-of-function studies without the limitations of single-cell clonal selection, preserving population-level heterogeneity relevant to cancer biology and ciliary research. The knockout model is generated using high-efficiency CRISPR/Cas9 ribonucleoprotein delivery, resulting in targeted disruption of DNAAF2 expression across the cell population. The polyclonal format is particularly suited for pooled functional genomics screens, drug-target validation, and studies where clonal variation could confound interpretation of DNAAF2-dependent phenotypes.
AGS cells, derived from a human gastric adenocarcinoma, serve as a widely used epithelial model for gastric mucosal biology, Helicobacter pylori infection, and gastric carcinogenesis. These adherent cells exhibit typical epithelial morphology and retain key signaling networks involved in gastric epithelial homeostasis, making them a relevant host for investigating genes linked to ciliary function and cancer. Although AGS cells are not classically ciliated under standard culture conditions, they can form primary cilia upon serum starvation or specific induction protocols, allowing conditional investigation of ciliary assembly and motility-associated factors. This host background also provides a platform to explore non-ciliary functions of DNAAF2 within gastric epithelial transformation and tumor progression.
DNAAF2 (dynein axonemal assembly factor 2) encodes a cytoplasmic protein essential for the preassembly of axonemal dynein arms, which are multi-subunit motor complexes required for ciliary beat generation. The protein functions downstream of master ciliogenic transcription factors RFX and FOXJ1, interacting with a conserved set of assembly cofactors including DNAAF1, LRRC6, and SPAG1. These interactions facilitate the folding and stabilization of dynein heavy chains before their transport and docking onto axonemal microtubules. Disruption of DNAAF2 abrogates both outer and inner dynein arm assembly, leading to immotile cilia and impaired mucociliary clearance. In humans, loss-of-function mutations in DNAAF2 are causally linked to primary ciliary dyskinesia (PCD) and Kartagener syndrome, characterized by chronic respiratory infections, situs inversus, and infertility. The DNAAF2-dependent pathway represents a critical node in cytoplasmic dynein arm assembly, integrating signals from upstream ciliogenesis programs and delivering functional dynein complexes to the ciliary compartment.
In the AGS cellular context, DNAAF2 knockout provides a unique model to dissect both canonical and potentially non-canonical roles of this assembly factor. While AGS cells do not constitutively display robust motile cilia, they express basal levels of ciliogenic machinery and can be induced to form primary cilia, enabling the study of DNAAF2??s involvement in early ciliogenesis and ciliary maintenance. Moreover, emerging evidence suggests that ciliary proteins may contribute to cancer-relevant processes independently of cilia, including cell cycle regulation, migration, and signaling pathway modulation. The DNAAF2 knockout AGS polyclonal population thus allows researchers to investigate whether loss of DNAAF2 alters gastric epithelial cell proliferation, migration, or response to oncogenic stimuli, potentially uncovering novel functions beyond its established role in dynein assembly.
Key research applications include quantitative assessment of DNAAF2 mRNA and protein knockdown by RT-qPCR and western blotting, immunofluorescence analysis of dynein arm components and ciliary markers following ciliogenesis induction, and functional assays such as cell proliferation and migration. The model is also suitable for drug screening aiming to restore or bypass DNAAF2 function, as well as for studying interactions with known cofactors (DNAAF1, LRRC6, SPAG1) in gastric epithelial cells. Researchers investigating primary ciliary dyskinesia, motile ciliopathies, or the emerging links between ciliary genes and gastric cancer will find this knockout population a valuable tool. For additional technical details or ordering information, please contact Ascent Research.
