The GPR171 Knockout PaTu 8988t Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population derived from the human pancreatic ductal adenocarcinoma cell line PaTu 8988t, featuring targeted disruption of the GPR171 locus. GPR171 encodes a G protein-coupled receptor that is endogenously activated by the proSAAS-derived neuropeptide BigLEN. The polyclonal format preserves genetic heterogeneity, minimizing clonal artifacts and enabling robust loss-of-function analyses.
PaTu 8988t is a metastatic pancreatic cancer line isolated from a liver metastasis, characterized by epithelial morphology and an oncogenic KRAS mutation, representing a model of aggressive PDAC. This KRAS-driven background renders the cells dependent on constitutively active MAPK/ERK and PI3K/AKT signaling, providing a relevant context for studying how GPR171 integrates into existing oncogenic pathways and influences tumor cell behavior.
At the molecular level, GPR171 couples to G??i/o proteins, inhibiting adenylyl cyclase and reducing cAMP production, which in turn downregulates PKA activity and phosphorylation of the transcription factor CREB. Additionally, receptor activation triggers ??-arrestin-mediated signaling that can stimulate the MAPK/ERK cascade, leading to ERK1/2 phosphorylation and downstream transcriptional reprogramming that modulates cell proliferation and migration. These dual pathways position GPR171 as a modulator of both metabolic and proliferative signals, with potential implications for cancer cell growth and motility.
Knockout of GPR171 in PaTu 8988t cells allows dissection of BigLEN-dependent signaling within a KRAS-mutant milieu. The loss of GPR171 may attenuate cAMP/PKA and ERK pathway modulation, offering a means to investigate the receptor??s contribution to pancreatic cancer hallmarks such as uncontrolled proliferation and enhanced migration. This model thus facilitates the study of crosstalk between metabolic GPCRs and core oncogenic drives.
These polyclonal knockout cells are well-suited for a suite of applications, including gene expression analysis by RT-qPCR and western blotting, functional assays for proliferation, migration, and colony formation, and pathway-specific measurements such as cAMP accumulation and phospho-ERK detection. These cells are also amenable to pooled genetic screens and high-throughput drug testing. The model supports pancreatic cancer research, GPCR signal transduction studies, and drug target validation. For further information, please contact Ascent Research.
