The GPR171 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the GPR171 gene in the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool of edited cells provides a versatile loss-of-function model for investigating GPR171-dependent signaling without the bias of single-cell clonal selection. The polyclonal format captures a broad spectrum of genomic modifications, ensuring robust representation of the knockout phenotype for downstream functional studies.
The HT29 cell line was originally isolated from a primary colorectal adenocarcinoma of a 44-year-old female patient. These cells exhibit an adherent epithelial morphology and are widely utilized as a model system for colorectal cancer biology. HT29 cells retain key features of intestinal epithelium, including the capacity for differentiation and expression of enterocytic markers, making them highly relevant for studies of tumor cell signaling, metabolism, and therapeutic responses in a colorectal cancer context.
GPR171 encodes an orphan G protein-coupled receptor that serves as a high-affinity receptor for the proSAAS-derived neuropeptide BigLEN. Upon ligand binding, GPR171 primarily couples to G??i/o proteins, leading to inhibition of adenylyl cyclase and a consequent reduction in intracellular cyclic AMP (cAMP) levels. This attenuates protein kinase A (PKA) activity and modulates downstream phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and the cAMP response element-binding protein (CREB). GPR171 signaling also involves ??-arrestin2 recruitment and exhibits crosstalk with the ??-opioid receptor in feeding circuits. Disruption of GPR171 abrogates BigLEN-induced Gi/o signaling, resulting in elevated cAMP and altered ERK1/2 phosphorylation, which may impact metabolic and proliferative pathways in colorectal cancer cells.
In the HT29 colorectal cancer background, GPR171 knockout provides a disease-relevant platform to dissect the receptor??s contributions to tumor cell metabolism and growth. Because HT29 cells are of intestinal epithelial origin, they express components of nutrient-sensing and energy homeostasis networks that intersect with GPR171. Loss of GPR171 function may perturb neuropeptide-driven metabolic regulation, potentially affecting pathways that control cell proliferation, survival, and metabolic reprogramming??hallmarks of colorectal carcinogenesis. This model thus enables researchers to examine how BigLEN-GPR171 signaling interfaces with oncogenic cascades and whether its disruption alters tumorigenic properties in vitro and in vivo.
These polyclonal knockout cells are suitable for a wide range of research applications, including functional characterization of GPR171 in colorectal cancer, high-throughput GPCR drug screening, and target validation for obesity and metabolic syndrome. Researchers can employ cAMP accumulation assays to verify loss of Gi/o coupling, phospho-ERK western blotting to probe mitogenic signaling, RT-qPCR to quantify downstream transcripts such as NPY and AgRP, and proliferation or colony formation assays to assess growth phenotypes. The cells are also compatible with xenograft tumor models and metabolic flux analyses to explore the receptor??s role in cancer metabolism and energy homeostasis. For further information or technical assistance, please contact Ascent Research.
