The GPR171 Knockout HCT 116 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, engineered to disrupt the GPR171 gene and establish a loss-of-function model. This polyclonal knockout pool, obtained through CRISPR/Cas9-mediated gene disruption, enables researchers to interrogate the functional consequences of GPR171 deficiency in a heterogeneous cellular context, avoiding the clonal artifacts that can arise from single-cell-derived lines. The targeted disruption of GPR171 provides a versatile tool for studying orphan GPCR biology and its implications in cancer cell signaling, immune modulation, and therapeutic target validation.
The HCT 116 host cell line is a widely characterized epithelial colorectal carcinoma model exhibiting microsatellite instability and a well-documented KRAS mutation, making it particularly relevant for investigating tumor cell proliferation, apoptosis, and drug response mechanisms. This genetic background, combined with the knockout of GPR171, offers a robust platform for dissecting signaling pathways that contribute to colorectal cancer progression and resistance, while maintaining the endogenous genomic architecture of a clinically pertinent tumor model.
GPR171 is an orphan G protein-coupled receptor that predominantly couples to G??i/o proteins, leading to inhibition of adenylyl cyclase and a consequent decrease in intracellular cAMP levels. This signaling cascade propagates through downstream effectors including PKA and CREB, and intersects with key oncogenic pathways such as MAPK/ERK and PI3K/AKT. Additionally, GPR171 interacts with ??-arrestins, which can mediate receptor desensitization and G protein-independent signaling. By disrupting GPR171, the polyclonal knockout cells permit the examination of altered signal transduction dynamics across these interconnected networks, shedding light on the receptor’s role in modulating cell growth, survival, and immune regulatory functions.
In the context of HCT 116 colorectal carcinoma cells, the loss of GPR171 expression may significantly impact cellular responses to microenvironmental cues and therapeutic agents. Given the putative involvement of GPR171 in immune regulation and its expression in intestinal tissues, this knockout model is particularly valuable for elucidating mechanisms of immune evasion and inflammation-associated carcinogenesis. Researchers can use these cells to compare proliferative capacity, apoptotic thresholds, and signaling pathway activation profiles with wild-type HCT 116 cells, thereby identifying GPR171-dependent phenotypes that may inform drug targeting strategies in colorectal cancer and inflammatory bowel disease.
Typical research applications for the GPR171 Knockout HCT 116 Polyclonal Cells include functional assays such as cAMP measurement to assess adenylyl cyclase activity, ERK phosphorylation detection via Western blot or flow cytometry, and cell proliferation or apoptosis assays to evaluate growth and survival. The polyclonal nature of the knockout population makes it suitable for CRISPR knockout validation by sequencing or Western blot, as well as for transcriptomic analyses like RNA-seq to identify global gene expression changes. These cells are also well-suited for drug target screening campaigns focused on orphan GPCRs and immune evasion mechanisms. For further details or technical inquiries, please contact Ascent Research.
