The ASAP3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. The ASAP3 gene has been disrupted to generate a loss-of-function model, eliminating functional protein expression. The polyclonal format provides a heterogeneous pool of edited cells, suitable for studying ASAP3-dependent processes without clonal selection artifacts. This product is intended for advanced biomedical research into GTPase regulation, cell migration, and cancer metastasis.
The HT29 parental line is an adherent epithelial cell model originating from a human colorectal adenocarcinoma. It retains differentiated intestinal features such as epithelial barrier properties and transporter expression, making it a well-established system for colorectal cancer biology, drug transport studies, and mucosal research. Its relevance to intestinal physiology and pathology provides a physiologically meaningful context for investigating molecular drivers of tumor progression.
ASAP3 functions as a GTPase-activating protein for Arf1 and Arf6, catalyzing the hydrolysis of GTP to downregulate Arf-mediated actin cytoskeleton remodeling and membrane trafficking. Its activity is modulated by upstream signals from integrin adhesion receptors, receptor tyrosine kinases, phospholipase D, and the PI3K pathway. ASAP3 interacts directly with focal adhesion and cytoskeletal components such as cortactin, paxillin, and SH3 domain-containing adaptors, and its downstream effects converge on the regulation of focal adhesion kinase (FAK), paxillin phosphorylation, and actin dynamics, ultimately controlling cell migration and invasion.
Knockout of ASAP3 in the colorectal adenocarcinoma background is predicted to elevate Arf-GTP levels, particularly Arf6-GTP, leading to increased actin polymerization, enhanced focal adhesion turnover, and more dynamic membrane protrusions. These molecular changes are expected to amplify invasive and migratory behavior, making this polyclonal knockout population a relevant tool for dissecting the mechanisms that drive colorectal cancer metastasis. The model enables the study of how ASAP3-dependent Arf regulation intersects with integrin-mediated adhesion and motility pathways in a tumor cell context.
Researchers can apply these cells in a variety of functional and biochemical assays, including transwell migration and invasion, wound healing, and cell adhesion measurements, to directly probe the contribution of ASAP3 to metastatic traits. Complementary techniques such as Arf6-GTP pull-downs, phospho-signaling analysis, and co-immunoprecipitation of interaction partners like FAK, paxillin, and cortactin allow detailed mapping of signaling network perturbations. Immunofluorescence for actin filaments and focal adhesion components further enables visualization of cytoskeletal remodeling. For technical inquiries or order support, please contact Ascent Research.
