The AHNAK2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, designed for loss-of-function studies of the AHNAK2 scaffold protein. This polyclonal pool contains heterogeneous genetic edits that disrupt functional AHNAK2 expression, providing a robust model to investigate its roles in cell adhesion, migration, and tumor progression without the need for single-cell clonal isolation. The product enables researchers to examine AHNAK2-dependent phenotypes in a physiologically relevant epithelial background.
The parental HT29 cell line is an established model of human colorectal adenocarcinoma, originally isolated from a primary tumor. HT29 cells exhibit epithelial morphology and adherent growth, and they harbor a mutation in the APC tumor suppressor gene, leading to constitutive activation of the Wnt/??-catenin pathway. They also possess mutant p53, which contributes to genomic instability. These properties make HT29 cells highly suitable for investigating colorectal cancer mechanisms, including epithelial?Cmesenchymal transition, invasion, and response to differentiation agents, and they are commonly used in drug discovery and cancer biology research.
AHNAK2 encodes a large scaffold protein that organizes the actin cytoskeleton and modulates cell adhesion. It directly interacts with actin and the annexin A2/S100A10 complex to regulate actin dynamics and focal adhesion turnover. In colorectal cancer, AHNAK2 is transcriptionally activated by ??-catenin/TCF and TGF-??1/SMAD2/3 signaling, driving EMT. AHNAK2 also promotes activation of the MAPK/ERK pathway and AKT phosphorylation, which enhance cell migration and invasion. Additionally, AHNAK2 interacts with the related scaffold AHNAK, contributing to calcium signaling and cytoskeletal remodeling.
Given the constitutively active Wnt/??-catenin pathway in HT29 cells due to APC mutation, this AHNAK2 knockout model is particularly valuable for dissecting the specific contribution of AHNAK2 to ??-catenin-driven transcriptional programs and associated malignant phenotypes. Disruption of AHNAK2 in this context allows researchers to delineate its role in actin cytoskeletal reorganization, cell adhesion, and the acquisition of mesenchymal traits, such as vimentin upregulation and E-cadherin loss. The polyclonal knockout population provides a heterogeneous system that more closely mimics the genetic variability found in tumors, enhancing the translational relevance of findings.
The AHNAK2 Knockout HT29 Polyclonal Cells are ideally suited for a range of mechanistic and functional studies. Researchers can employ Western blot and RT-qPCR to validate AHNAK2 disruption and assess downstream targets, while migration and invasion assays quantify the impact on metastatic potential. Co-immunoprecipitation and immunofluorescence enable investigation of interactions with actin, annexin A2, and S100A10. Phospho-signaling arrays and RNA-seq can map AHNAK2-dependent signaling networks and transcriptomic changes. These cells also support drug target validation and high-content screening for modulators of AHNAK2-driven pathways. For further technical details or to discuss your specific experimental requirements, please contact Ascent Research.
