The AKAP12 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma cell line HT29. In this model, the AKAP12 gene, which encodes the A-kinase anchoring protein 12 scaffold protein (also known as Gravin), has been disrupted to generate a heterogeneous pool of cells with loss-of-function mutations. This product provides researchers with a robust tool for dissecting the tumor-suppressive roles of AKAP12 without the biases inherent in clonal selection.
HT29 cells originated from a primary colon adenocarcinoma and exhibit characteristic features of malignant colonic epithelium, including mutations in oncogenes and tumor suppressors such as BRAF (V600E), TP53, and PIK3CA. They grow as adherent monolayers, form tight junctions, and maintain the capacity for polarization, making them a well-established model for colorectal cancer biology, epithelial barrier function, and tumor cell migration.
AKAP12 functions as a multivalent scaffold protein that orchestrates cAMP/PKA and PKC signaling by anchoring PKA-RII subunits and PKC to the actin cytoskeleton. This scaffold facilitates the phosphorylation of downstream targets such as cofilin, myosin light chain, vinculin, and paxillin, thereby regulating actin dynamics, focal adhesion turnover, and cell adhesion. AKAP12 interacts directly with F-actin, E-cadherin, and ??-catenin, linking cell?Ccell adhesion complexes to the cytoskeleton. Upstream, AKAP12 is regulated by cAMP produced through ??-adrenergic receptor-activated adenylyl cyclase and by Src kinase-mediated tyrosine phosphorylation, while epigenetic silencing via DNA methylation and histone deacetylation often reduces its expression in cancers.
In the HT29 background, disruption of AKAP12 is expected to relieve its tumor-suppressive constraints, leading to enhanced cell migration, invasion, and proliferation. Loss of AKAP12-mediated scaffolding may destabilize adherens junctions and focal adhesions, promote actin reorganization, and activate signaling pathways that drive epithelial-to-mesenchymal transition. These changes mirror aggressive colorectal cancer phenotypes and provide a relevant model for studying the contribution of AKAP12 to tumor progression and metastasis.
This knockout cell population is ideally suited for investigating mechanisms of colorectal cancer metastasis, cell migration, and barrier disruption using functional assays such as wound healing and Transwell invasion. It also enables exploration of cAMP-dependent signaling dynamics and tumor suppressor pathways through western blotting for AKAP12 and phospho-PKA substrates, RT?qPCR, and immunofluorescence for F-actin or vinculin. Additional applications include drug sensitivity screening and apoptosis or proliferation assays to assess therapeutic responses. For technical inquiries or ordering information, please contact Ascent Research.
