GPR39 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal adenocarcinoma cell line. This loss-of-function model carries a targeted disruption of the GPR39 gene, eliminating expression of the zinc-sensing G protein-coupled receptor. The polyclonal format provides a heterogeneous knockout pool suitable for pooled functional screens and robust population-level phenotypic analyses. Rigorous quality control confirms the absence of GPR39 protein, enabling precise interrogation of receptor-mediated signaling cascades in a genetically defined cancer cell background.
The HCT 116 host cell line is a widely utilized model of colorectal carcinoma, characterized by a KRAS G13D mutation, ??-catenin (CTNNB1) mutation, microsatellite instability, and wild-type p53. This genetic profile renders the cells highly relevant for studying oncogenic signaling networks and tumor suppressor pathways. The epithelial origin and adherent growth properties facilitate a broad range of in vitro assays. The addition of GPR39 knockout extends the utility of this line to dissect zinc-sensing receptor functions in the context of deregulated MAPK and Wnt/??-catenin signaling.
GPR39 functions as a zinc-activated receptor that couples predominantly to G??q and G??s heterotrimeric G proteins. Upon Zn2? binding, it triggers phospholipase C?? (PLC??)-mediated production of inositol 1,4,5-trisphosphate (IP?) and diacylglycerol, leading to intracellular calcium mobilization and protein kinase C activation. Concurrently, Gs stimulates adenylyl cyclase (AC) to elevate cAMP levels and activate protein kinase A (PKA). The receptor also engages ??-arrestin-1/2, which scaffolds and sustains MAPK/ERK and PI3K/AKT signaling modules. Key downstream effectors include phosphorylated ERK1/2 (MAPK3/1) and AKT (AKT1/2/3), as well as transcription factors CREB, Jun, and Fos, thereby promoting cell cycle progression and anti-apoptotic programs through modulation of Bcl-2 family members. The receptor interacts with scaffold proteins such as NHERF1/PDZK1, which may influence its trafficking and signal compartmentalization.
In the HCT 116 colorectal cancer model, GPR39 signaling converges on proliferative and survival pathways frequently hyperactivated by oncogenic KRAS and ??-catenin mutations. Ablation of GPR39 enables direct assessment of receptor contributions to zinc-dependent growth stimulation, cytosolic Ca2? dynamics, and transcriptional regulation. This model helps distinguish GPR39-specific effects from other zinc-responsive pathways and provides a clean background for structure?Cfunction studies and pharmacological profiling of GPR39 agonists or antagonists. Given the receptor??s emerging roles in apoptosis resistance and metabolic adaptation, the knockout cells serve as a critical tool for studying tumor cell fitness under stress conditions.
Typical research applications include colorectal cancer signaling, zinc biology, and GPCR drug target discovery. The cells are suitable for proliferation assays (MTT, BrdU incorporation, colony formation), apoptosis studies (Annexin V staining), intracellular calcium flux analyses, cAMP ELISAs, and phospho-ERK/AKT immunoblotting. Transcriptomic profiling by RNA-seq or targeted gene expression analysis (RT-qPCR) can reveal GPR39-dependent gene networks. This polyclonal knockout population is particularly valuable for high-content screening and pathway dissection where clonal variation is not desired. For further information or customized orders, please contact Ascent Research.
