The IFT88 Knockout AGS Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, in which the gene encoding intraflagellar transport 88 (IFT88) has been functionally disrupted. This polyclonal pool consists of a heterogeneous mixture of edited cells collectively lacking wild-type IFT88 expression, providing a loss-of-function model suitable for studying the roles of primary cilia and intraflagellar transport in gastric epithelial biology. The use of a polyclonal population, rather than a clonal line, captures the diversity of editing outcomes while maintaining robust knockout efficiency, enabling experiments that interrogate the overall impact of IFT88 ablation without clonal bias.
The AGS cell line is a widely used model of gastric adenocarcinoma, representing epithelial cells from a stomach tumor. These cells exhibit adherent growth and retain features of gastric epithelial differentiation, making them relevant for investigations of gastric cancer progression, mucosal homeostasis, and signaling pathway deregulation. The gastric epithelial context is particularly important given the emerging roles of primary cilia in gastrointestinal tissues and the association of ciliary defects with hyperproliferation and metaplasia. As an adherent human epithelial line, AGS cells offer a tractable system for genetic manipulation and phenotypic analysis, supporting high-resolution microscopy, biochemical assays, and functional screens.
IFT88 functions as a core subunit of the intraflagellar transport complex B (IFT-B), which mediates anterograde movement of cargo along the ciliary axoneme. It is essential for the assembly and maintenance of primary cilia, organelles that coordinate multiple developmental and homeostatic signaling pathways. In AGS cells, IFT88 knockout prevents cilium formation, leading to disruption of canonical Hedgehog signaling??whereby GLI transcription factors (GLI1, GLI2, GLI3) are not properly processed??and aberrant WNT/??-catenin (CTNNB1) signaling. IFT88 interacts directly with other IFT-B components (IFT20, IFT52, IFT57, IFT80, IFT172) and the kinesin-2 motor (KIF3A/KIF3B), and its function is required for the ciliary localization of the BBSome (BBS4, BBS5) and smoothened (SMO) receptor. Consequently, the signaling cascade involving SHH, PTCH1, SMO, SUFU, and the GLIs is attenuated, while WNT pathway components (WNT3A, FZD, DVL2, AXIN1, CTNNB1) exhibit altered activity. Upstream regulators such as RFX transcription factors (RFX3), FOXJ1, and HNF1B orchestrate IFT88 expression in response to cellular cues like serum starvation, further linking ciliogenesis to cell cycle control.
In the AGS gastric cancer background, loss of IFT88 and primary cilia provides a powerful tool to dissect how ciliary signaling influences epithelial cell proliferation, migration, and differentiation. The model is particularly relevant for studying the proposed tumor-suppressive roles of primary cilia in gastric mucosa and for exploring how ciliary dysfunction contributes to carcinogenesis. Additionally, this polyclonal knockout population enables investigation of ciliopathy-related mechanisms, as IFT88 mutations are associated with short-rib polydactyly syndrome and polycystic kidney disease, and ciliary defects are linked to retinal degeneration and other ciliopathies. By comparing wild-type and IFT88-null AGS cells, researchers can uncover context-specific functions of primary cilia in gastric epithelial homeostasis and disease.
Researchers can employ these IFT88 Knockout AGS Polyclonal Cells in a wide array of experiments, including immunofluorescence staining for acetylated tubulin and ARL13B to assess ciliary frequency and morphology, western blotting to measure changes in GLI1 and phospho-??-catenin levels, and RT-qPCR profiling of hedgehog target genes such as GLI1 and PTCH1. The cells are also compatible with functional assays evaluating cell migration and proliferation, enabling comprehensive analysis of ciliary influence on gastric cancer cell behavior. For additional details or personalized guidance on integrating this model into your research, please contact Ascent Research.
