The DYNC2H1 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, with targeted disruption of the DYNC2H1 gene. This population consists of cells harboring diverse editing events that collectively abolish DYNC2H1 protein expression, providing a robust loss-of-function model without clonal biases. The polyclonal format is particularly suited for population-level functional studies and reduces artifacts associated with single-clone expansion.
The AGS cell line is a well-established epithelial model derived from a human gastric adenocarcinoma, extensively used for investigating gastric physiology, Helicobacter pylori pathogenesis, and gastric cancer biology. AGS cells retain epithelial characteristics, including the ability to form polarized monolayers with tight junctions, and are competent for primary cilia formation, making them a relevant host for studying cilia-dependent signaling in the gastric epithelium.
DYNC2H1 encodes the heavy chain of the cytoplasmic dynein-2 motor, which powers retrograde intraflagellar transport (IFT) within primary cilia. As part of the dynein-2 complex, DYNC2H1 interacts with light intermediate chain DYNC2LI1 and adaptors WDR34 and WDR60, and cooperates with the IFT-A and IFT-B complexes to return signaling receptors and IFT particles from the ciliary tip to the cell body. Its expression is regulated by ciliogenic transcription factors FOXJ1 and RFX. DYNC2H1 function is critical for Hedgehog signal transduction, as retrograde IFT enables the processing of GLI transcription factors downstream of the receptor SMO and its regulator PTCH1, leading to expression of target genes such as CCND1. Knockout of DYNC2H1 thus impairs ciliary assembly and abrogates GLI-mediated transcriptional responses.
In the AGS gastric cancer context, DYNC2H1 disruption attenuates primary cilium formation and Hedgehog pathway activity, potentially altering cellular proliferation, migration, and differentiation. Given the emerging role of ciliary signaling in gastrointestinal malignancies, this polyclonal knockout model provides a valuable tool to dissect cilia-dependent contributions to gastric adenocarcinoma phenotypes without the confounding effects of clonal selection.
This toolset is suited for ciliopathy disease modeling, Hedgehog pathway analysis, and functional genomics in gastric cancer. Ciliary phenotypes can be examined by immunofluorescence for ARL13B and acetylated ??-tubulin, while protein-level changes in DYNC2H1 and GLI isoforms are assessed by western blot. Transcriptional consequences are measured via RT-qPCR for GLI targets, and functional impacts on cell migration and proliferation are evaluated using wound healing and MTT assays. For further assistance or to discuss customized applications, please contact Ascent Research.
