The GPRC5C Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal gene knockout population derived from HT29 human colorectal adenocarcinoma cells, designed to disrupt the GPRC5C locus. This heterogeneous pool of cells carries loss-of-function mutations in GPRC5C, abolishing orphan GPCR signaling and providing a robust system to dissect retinoic acid-induced growth suppression and apoptotic pathways. As a polyclonal product, it avoids clonal bias and supports bulk functional genomic studies in colorectal cancer research.
The parental HT29 cell line, established from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female, displays an epithelial phenotype and is widely used in intestinal biology, drug absorption, and oncology investigations. HT29 cells retain characteristics of differentiated intestinal epithelium, including polarized monolayer formation and brush-border enzyme expression, making them a physiologically relevant model for examining retinoic acid signaling in a malignant context.
GPRC5C is an orphan class C G protein-coupled receptor that transduces all-trans retinoic acid signals via coupling to G??q/11 and G?¦? subunits. Upon retinoic acid-bound RAR/RXR activation, GPRC5C modulates the MAPK/ERK cascade, reducing phosphorylated ERK (p-ERK) and shifting the balance of Bcl-2 family members toward pro-apoptotic Bax, leading to caspase-3-mediated apoptosis. Concurrently, GPRC5C downregulates cyclin D1, contributing to cell cycle arrest. This pathway integrates retinoic acid sensing with tumor-suppressive outcomes.
In the HT29 colorectal cancer model, GPRC5C knockout abrogates retinoic acid-induced growth suppression and apoptosis, mirroring loss-of-function states observed in aggressive malignancies with diminished retinoic acid sensitivity. The polyclonal knockout population recapitulates tumor heterogeneity, enabling assessment of GPRC5C deficiency on proliferation, survival, and metastatic potential. These cells are instrumental for studying GPCR-mediated signaling in colorectal tumorigenesis and for screening strategies to restore retinoic acid responsiveness.
Key applications include dissecting GPRC5C roles in retinoic acid signaling and MAPK/ERK regulation, validating GPRC5C as a therapeutic target, and evaluating differentiation therapies. Typical assays employ Western blotting for p-ERK, Bcl-2, Bax, and caspase-3; RT-qPCR for downstream targets; MTT proliferation; TUNEL apoptosis; and transwell migration/invasion. The polyclonal format facilitates pooled CRISPR screening and compound profiling. For further details or customized applications, please contact Ascent Research.
