The CASR Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with disruption of the calcium-sensing receptor (CASR) gene in the HCT 116 human colorectal carcinoma cell line. This knockout model is generated using CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous pool of CASR loss-of-function cells, suitable for studying gene function without clonal selection bias.
HCT 116 is a well-characterized colorectal carcinoma line with a KRAS G13D mutation, wild-type TP53, and microsatellite instability (MSI-H). Its epithelial morphology and high tumorigenicity make it a standard model for cancer research, particularly for dissecting oncogenic signaling and tumor suppressor mechanisms. The intact p53 and MSI-H status provide a defined genetic context for investigating genes like CASR.
CASR encodes a class C GPCR that senses extracellular Ca2?, Mg2?, L-amino acids, polyamines, and calcimimetics. Upon activation, it couples to G??q and G??i/o, stimulating PLC/IP3/Ca2? signaling while reducing cAMP. Downstream, it regulates MAPK/ERK, PI3K/AKT, and NF-??B pathways. CASR interacts with ??-arrestins, filamin A, and caveolin, and suppresses Wnt/??-catenin by promoting ??-catenin degradation. These pathways mediate diverse cellular responses including changes in gene transcription via AP-1 and regulation of parathyroid hormone (PTH) secretion. These molecular interactions position CASR as a key modulator of proliferation and differentiation.
In colorectal cancer, CASR is proposed to act as a tumor suppressor. Disrupting CASR in HCT 116 cells allows investigation of its role in ??-catenin stability, ERK activation, and calcium homeostasis. Loss of CASR in HCT 116 may mimic aspects of colorectal cancer progression, enabling assessment of its impact on cell proliferation, colony formation, and invasive capacity. This knockout model facilitates examination of CASR-mediated control of apoptosis, migration, and differentiation, providing insights into its contribution to colorectal carcinogenesis and potential as a therapeutic target.
Applications include Western blotting and RT-qPCR for downstream targets like ERK1/2 and ??-catenin, calcium imaging, proliferation and apoptosis assays, and Wnt/??-catenin reporter assays. Typical experiments also employ flow cytometry for apoptosis detection and migration/invasion assays to evaluate metastatic potential. The cells support drug screening for calcimimetics and calciolytics and study of CASR-dependent signaling networks. For further information, contact Ascent Research.
