The GAN Knockout AGS Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, engineered to disrupt the expression of GAN (gigaxonin). This loss-of-function model enables targeted investigation of gigaxonin-dependent ubiquitination and its role in cytoskeletal protein turnover within an epithelial gastric cancer background. The polyclonal format ensures a heterogeneous knockout population suitable for studying pooled genetic effects, drug responses, and signaling dynamics without the limitations of single-clone artifacts. As a tool for functional genomics, this product facilitates dissection of the CUL3-RING E3 ligase pathway and downstream cellular processes relevant to oncology and neurobiology.
The AGS cell line is a well-characterized adherent epithelial model derived from a human gastric adenocarcinoma. It is extensively employed in Helicobacter pylori pathogenesis studies, cancer biology research, and epithelial cell function assays. AGS cells exhibit typical epithelial morphology and harbor molecular features pertinent to gastric carcinogenesis, including dysregulated signaling networks. Their robustness in culture and compatibility with advanced imaging, cytotoxicity, and migration assays make them an ideal host for gene disruption studies focused on tumor cell biology and drug sensitivity profiling.
GAN encodes gigaxonin, a substrate adaptor protein that forms a CUL3-RING E3 ubiquitin ligase complex together with CUL3 and RBX1. This E3 ligase is activated by CUL3 neddylation involving NEDD8 and NAE1, and it specifically ubiquitinates key cytoskeletal substrates such as vimentin (VIM), neurofilament light chain (NEFL), peripherin (PRPH), GFAP, and ??III-tubulin (TUBB3). Ubiquitinated substrates are subsequently degraded by the 26S proteasome. Gigaxonin-mediated proteasomal degradation is therefore a critical mechanism for regulating intermediate filament turnover and maintaining cytoskeletal organization. Loss of GAN disrupts this pathway, leading to aberrant accumulation of intermediate filaments and altered cytoskeletal dynamics, which can influence cell morphology, adhesion, and migration.
In the AGS gastric cancer background, GAN knockout models the effects of impaired ubiquitin-dependent proteolysis on epithelial tumor cell behavior. Defective gigaxonin function causes accumulation of vimentin and other cytoskeletal elements, potentially altering epithelial-mesenchymal transition markers, migration, and invasion capabilities. This model is particularly relevant for studying how cytoskeletal protein homeostasis influences gastric adenocarcinoma progression and response to proteasome inhibitors or other targeted therapies. Moreover, the interplay between autophagy and the ubiquitin-proteasome system can be explored, as GAN deficiency may trigger compensatory autophagy or render cells more susceptible to proteotoxic stress.
Typical applications include quantitative immunoblotting and RT-qPCR to confirm GAN disruption, immunofluorescence or flow cytometry to assess vimentin and neurofilament accumulation, wound healing and transwell invasion assays to evaluate migration and invasion, and co-immunoprecipitation with CUL3 to probe complex integrity. Additionally, this model supports proteasome activity assays, ubiquitination analyses, and dose-response studies with proteasome inhibitors or autophagy modulators. It also serves as a tool for investigating giant axonal neuropathy mechanisms within an epithelial context. For further technical details, please contact Ascent Research.
