The HOMER2 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HT29 cells harboring targeted disruption of the human HOMER2 gene. This mixed knockout pool provides a genetically heterogeneous loss-of-function model, enabling investigation of HOMER2-dependent mechanisms without clonal selection bias. The polyclonal format preserves cellular diversity while ensuring widespread gene editing, making it suitable for bulk population studies where heterogeneous knockout effects reflect the complexity of biological systems.
HT29 cells are a well-characterized human colorectal adenocarcinoma line derived from a primary tumor, retaining epithelial morphology and the capacity for enterocytic differentiation under metabolic stress conditions. These cells serve as a widely accepted model for intestinal epithelial biology, colorectal cancer progression, and nutrient-dependent signaling. Their adherent growth and stable karyotype facilitate reproducible CRISPR/Cas9 editing and downstream phenotypic assays.
HOMER2 serves as an adaptor scaffold that organizes macromolecular complexes at the intersection of calcium and immune signaling. It interacts with mGluR1/5, IP3 receptors, TRPC channels, and SHANK scaffolds to control calcium flux and downstream effector coupling. A key function is the competitive regulation of NFAT: HOMER2 binds NFAT proteins in the cytoplasm, maintaining their phosphorylated state, whereas calcium-activated calcineurin dephosphorylates NFAT to promote nuclear translocation and transcriptional activity. The canonical pathway proceeds from T-cell receptor/CD3 complex activation through PLC??, IP3-dependent calcium release, calcineurin engagement, and NFAT-mediated expression of targets such as IL-2. Additional upstream regulators include neuronal activity and calcium influx through ORAI1 channels, placing HOMER2 at a convergence point for diverse stimuli.
In the HT29 colorectal cancer context, HOMER2 knockout is expected to perturb calcium?CNFAT signaling, potentially influencing cell proliferation, differentiation, and migration??processes critical to tumor progression and epithelial homeostasis. Given HOMER2??s association with autosomal dominant nonsyndromic hearing loss and immune disorders, this model also supports cross-disciplinary studies into the scaffolding protein??s role in non-neuronal tissues. The loss of HOMER2 may alter sensitivity to calcium-dependent stimuli, providing a platform for dissecting oncogenic signaling networks and evaluating therapeutic targets in colorectal adenocarcinoma.
Researchers can apply this polyclonal knockout model to study HOMER2??s function in colorectal cancer signaling, validate its role in NFAT pathway regulation, and screen for modulators of calcium-dependent processes in intestinal epithelia. Representative assays include Western blotting and RT-qPCR for knockout confirmation, calcium imaging with Fluo-4 AM, NFAT-luciferase reporter assays, EdU proliferation assays, transwell migration studies, and apoptosis analysis. For further information or technical support, please contact Ascent Research.
