We present the HPCAL1 Knockout HT29 Polyclonal Cells, a CRISPR/Cas9-edited polyclonal cell population designed for loss-of-function studies of the human HPCAL1 gene. This product enables investigation of HPCAL1??s role in calcium-dependent signal transduction within an epithelial context. The knockout pool is generated by transient delivery of CRISPR/Cas9 ribonucleoproteins targeting HPCAL1, resulting in a heterogeneous mixture of gene-disrupted HT29 cells. The polyclonal format preserves biological variability and avoids clonal selection artifacts, making it suitable for population-level functional assays. Each lot is cryopreserved and quality-controlled to ensure viability and proliferative capacity.
The parental HT29 cell line is a widely employed model for colorectal adenocarcinoma, originally isolated from a primary tumor of a 44-year-old Caucasian female. These cells exhibit epithelial morphology and retain characteristic features of colon carcinoma, including aberrant Wnt signaling. HT29 cells are amenable to differentiation protocols and are frequently utilized to study intestinal epithelial biology, oncogenic transformation, and drug responses. Their genetic background provides a relevant system for interrogating colorectal cancer pathways influenced by HPCAL1 disruption.
HPCAL1 encodes a neuron-specific calcium-binding protein that belongs to the recoverin family. It functions as a calcium sensor, interacting with intracellular calcium ions, calmodulin, and the neuronal calcium sensor protein NCS1. HPCAL1 is regulated by calmodulin-dependent kinase, protein kinase C, and Wnt signaling, placing it at the intersection of multiple pathways. Downstream, HPCAL1 influences AMPA receptor trafficking, the MAPK/ERK cascade, and exocytosis machinery, potentially through its association with IL1RAPL1. Key representative components of its signaling network include CALM1, CAMK2, CTNNB1 (??-catenin), and TCF4. In HT29 cells, HPCAL1 disruption may perturb calcium-dependent modulation of these pathways, thereby affecting transcriptional programs governed by ??-catenin/TCF4 complexes and MAPK-driven processes.
Loss of HPCAL1 in HT29 cells provides a reductionist model to dissect its contributions to calcium-mediated signaling in colorectal cancer. Given that calcium signaling regulates proliferation, differentiation, and apoptosis, HPCAL1 knockout may alter these processes in the context of oncogenic Wnt pathway activation. The HT29 background, with its constitutive ??-catenin activity, offers a platform to study crosstalk between calcium-dependent modules and transcriptional outputs. Researchers can explore how HPCAL1 loss modifies chemosensitivity, invasive potential, or differentiation status, bridging gaps between neurodevelopmental gene function and epithelial tumor biology.
The HPCAL1 Knockout HT29 Polyclonal Cells are suited for diverse functional genomics applications. They enable quantitative analysis of gene expression changes via RT-qPCR and RNA-seq, and validation of protein-level disruption by Western blotting. Calcium imaging assays can directly assess alterations in intracellular calcium dynamics. Proliferation and viability can be measured using MTT assays, while flow cytometry facilitates cell cycle and apoptosis profiling. These cells are valuable for drug screening studies targeting calcium or MAPK pathways in colorectal cancer models. For further information, please contact Ascent Research.
