The AGFG2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product introduces a loss-of-function mutation in the AGFG2 gene, which encodes an ARF GTPase-activating protein critical for endocytic trafficking. The polyclonal format provides a genetically heterogeneous knockout pool ideal for pooled functional assays without clonal selection.
HT29 cells originate from a 44-year-old female colorectal adenocarcinoma and are a well-established model for intestinal epithelial biology and colorectal cancer research. These adherent cells display characteristics relevant to studying oncogenic signaling, drug responses, and metastasis. Their defined genetic background supports reproducible investigation of gene function, and the AGFG2 knockout extends the model??s utility to dissect membrane trafficking roles in tumor cell behavior.
AGFG2 functions as a GTPase-activating protein for ARF GTPases, notably ARF1 and ARF5, promoting GTP hydrolysis to regulate COPI vesicle formation and endosomal recycling. It resides downstream of ARF activation and upstream of COPI subunits such as COPB1 and endosomal Rab proteins, while also interacting with the HIV-1 Gag protein. Through these interactions, AGFG2 modulates integrin recycling and endocytic sorting, influencing cell adhesion and migration. This signaling node links membrane dynamics to cellular responses, with regulatory inputs from unknown kinases potentially modulating its activity.
In HT29 cells, AGFG2 knockout disrupts endocytic trafficking and the recycling of adhesion molecules, which may impair cell migration and invasion??processes central to colorectal cancer metastasis. The model thus enables study of AGFG2-dependent mechanisms driving malignancy. Additionally, the known role of AGFG2 in HIV-1 replication via Gag interaction makes these cells a useful platform for investigating host factor contributions to viral assembly in a colonic epithelial context.
These polyclonal knockout cells are designed for diverse applications, including Western blotting and RT-qPCR for knockout validation, immunofluorescence to visualize endosomal markers, and migration/invasion assays to assess metastatic potential. Co-immunoprecipitation can probe interactions with ARF1, ARF5, or COPI components, while drug sensitivity studies may reveal AGFG2-related chemoresistance. The model is suitable for exploring endocytic trafficking, HIV-1 replication, and colorectal cancer progression. For additional information, contact Ascent Research.
